U.S. patent application number 14/027698 was filed with the patent office on 2014-01-16 for method for irradiating a sample with focused acoustic energy including a fully solid coupler.
This patent application is currently assigned to BIOCARTIS SA. The applicant listed for this patent is BIOCARTIS SA. Invention is credited to Contantijn W.M. Brantjes, Ronald De Gier, Michiel De Jong, Nicolaas B. Roozen, Sergei Shulepov, Louis Stroucken, Hendrick S. Van Damme, Marloes M.E.B. Van De Wal, Arie R. Van Doorn.
Application Number | 20140017694 14/027698 |
Document ID | / |
Family ID | 42235742 |
Filed Date | 2014-01-16 |
United States Patent
Application |
20140017694 |
Kind Code |
A1 |
Van Doorn; Arie R. ; et
al. |
January 16, 2014 |
METHOD FOR IRRADIATING A SAMPLE WITH FOCUSED ACOUSTIC ENERGY
INCLUDING A FULLY SOLID COUPLER
Abstract
A method for irradiating a sample with focused acoustic energy
is provided. The generated acoustic energy is transmitted from the
source to the sample via a completely dry propagation path. At
least two different focal regions are generated with the source for
generating acoustic energy.
Inventors: |
Van Doorn; Arie R.; (AE
Eindhoven, NL) ; De Gier; Ronald; (AE Eindhoven,
NL) ; Stroucken; Louis; (AE Eindhoven, NL) ;
Van De Wal; Marloes M.E.B.; (AE Eindhoven, NL) ;
Shulepov; Sergei; (AE Eindhoven, NL) ; Roozen;
Nicolaas B.; (AE Eindhoven, NL) ; Brantjes;
Contantijn W.M.; (AE Eindhoven, NL) ; Van Damme;
Hendrick S.; (AE Eindhoven, NL) ; De Jong;
Michiel; (AE Eindhoven, NL) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
BIOCARTIS SA |
LAUSANNE |
|
CH |
|
|
Assignee: |
BIOCARTIS SA
LAUSANNE
CH
|
Family ID: |
42235742 |
Appl. No.: |
14/027698 |
Filed: |
September 16, 2013 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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13273677 |
Oct 14, 2011 |
8563324 |
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14027698 |
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PCT/CH2010/000093 |
Apr 9, 2010 |
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13273677 |
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Current U.S.
Class: |
435/6.12 ;
435/173.7; 435/173.9; 435/29; 436/147; 436/149; 436/166;
436/174 |
Current CPC
Class: |
G01N 1/44 20130101; Y10T
436/25 20150115 |
Class at
Publication: |
435/6.12 ;
436/174; 435/173.9; 435/173.7; 436/166; 436/149; 436/147;
435/29 |
International
Class: |
G01N 1/44 20060101
G01N001/44 |
Foreign Application Data
Date |
Code |
Application Number |
Apr 14, 2009 |
EP |
09157850.0 |
Claims
1. A method of irradiating a sample with focused acoustic energy to
treat the sample, the method comprising the following steps:
providing a cartridge with a fully solid coupler; placing the
sample in a chamber of the cartridge; inserting the cartridge into
an instrument having a single source for generating acoustic energy
so that a completely dry coupling of the acoustic energy between
the source and the cartridge is provided, wherein the cartridge and
the instrument are separable; and generating at least two different
focal regions with the single source for generating acoustic
energy.
2. A method according to claim 1, wherein the step of generating
the at least two different focal regions includes one or more of
the following: using said source for generating acoustic energy and
a hybrid lens for focusing the acoustic energy onto the sample;
using said source for generating acoustic energy with different
roughness zones; exciting said source for generating acoustic
energy differently at different positions of said source; and any
combination thereof.
3. A method according to claim 2, wherein the lens is selected from
the group consisting of: a lens being a physically separate
component placed between the source and the cartridge, a lens being
part of the source, the source with a focusing shape being the
lens, an array of sources that yield to focused acoustic energy, a
lens being part of the cartridge, a lens made out of a polymer
having a low acoustic attenuation characteristic, a metal lens, a
ceramic lens, a polypropylene lens, an aluminum lens, a hybrid
lens, and any combination thereof.
4. A method according to claim 1, wherein said source for
generating acoustic energy is configured so that the acoustic
energy is high intensity focused ultra sound (HiFu).
5. A method according to claim 1, wherein said method further
comprises: inserting the cartridge into the instrument such that a
propagation path for transmitting the acoustic energy from the
source to the sample is formed, said propagation path consisting
only of non-fluidic matter.
6. A method according to claim 5, wherein the propagation path has
a gradient of an acoustic impedance that is monotonously decreasing
in a direction from the source to the sample.
7. A method according to claim 1, wherein the fully solid coupler
comprises a material selected from the group consisting of: solid
gel, rubber, elastic foil, polymer based material, thermoplastic
polymers, polymer having a low acoustic attenuation characteristic,
metal, semiconductor, ceramic, polypropylene, aluminum, and a stack
of these materials.
8. A method according to claim 1, wherein inserting the cartridge
into the instrument causes the fully solid coupler to be physically
contacted with an acoustic window of the cartridge, said acoustic
window made of a flexible material.
9. A method according to claim 8, wherein the fully solid coupler
has a first contact surface for contacting the acoustic window, the
cartridge has a second contact surface for contacting the acoustic
window, and at least one of the first contact surface, the second
contact surface and the acoustic window has a surface roughness
value selected from the group consisting of smaller than 0.5
micrometers, smaller than 1 micrometers, and smaller than 2
micrometers.
10. A method according to claim 1, wherein said method further
comprises: irradiating the sample in one single chamber of the
cartridge with focused acoustic energy to apply pretreatment and
lysis thereto, wherein the pretreatment is a method selected from
the group consisting of mixing with a reagent, circulation, release
of a cell, pathogen and matrix from a swab, release of a cell,
pathogen and matrix from a brush, liquefaction, incubation of the
sample with a reagent at room temperature or elevated temperature,
shaking, mixing; stifling, extraction, NA extraction, flow
generation, sample homogenation, separating by centrifuging, and
any combination thereof, and wherein lysis is a method selected
from the group consisting of mixing with a reagent, circulation,
lysis of microorganisms, incubation of the sample with a reagent at
room or elevated temperature, and any combination thereof.
11. A method according to claim 1, wherein the fully solid coupler
is made out of a polymer based material having a glass transition
temperature T.sub.g selected from the group consisting of:
T.sub.g.gtoreq.-30.degree. C.; T.sub.g.gtoreq.-10.degree. C.,
T.sub.g.gtoreq.-5.degree. C.; T.sub.g.gtoreq.20.degree. C.;
T.sub.g.gtoreq.40.degree. C.; T.sub.g.gtoreq.60.degree. C.;
T.sub.g.gtoreq.80.degree. C.; T.sub.g.gtoreq.100.degree. C.;
T.sub.g.gtoreq.120.degree. C.; T.sub.g.gtoreq.130.degree. C.;
T.sub.g.gtoreq.140.degree. C.; T.sub.g.gtoreq.150.degree. C.; and
T.sub.g.gtoreq.160.degree. C.
12. A method according to claim 11, wherein the polymer based
material has been cured at a curing temperature T.sub.c selected
from the group consisting of: T.sub.c.gtoreq.20.degree. C.;
T.sub.c.gtoreq.40.degree. C.; T.sub.c.gtoreq.60.degree. C.;
T.sub.c.gtoreq.70.degree. C.; T.sub.c.gtoreq.80.degree. C.;
T.sub.c.gtoreq.90.degree. C.; T.sub.c.gtoreq.100.degree. C.;
T.sub.c.gtoreq.110.degree. C.; T.sub.c.gtoreq.120.degree. C.;
T.sub.c.gtoreq.130.degree. C.; T.sub.c.gtoreq.140.degree. C.;
T.sub.c.gtoreq.150.degree. C.; T.sub.c.gtoreq.160.degree. C.;
T.sub.c.gtoreq.170.degree. C.; and T.sub.c.gtoreq.180.degree.
C.
13. A method according to claim 1, wherein said method further
comprises: applying at least one measurement to the sample selected
from the group consisting of: optical measurements, magnetic
measurements, thermal measurements, electrical measurements,
chemical measurements, sonic measurements, and any combination
thereof.
Description
RELATED APPLICATIONS
[0001] This application is a divisional of U.S. application Ser.
No. 13/273,677, filed Oct. 14, 2011, which is a continuation of
International Application No. PCT/CH2010/000093, filed Apr. 9,
2010, which claims the benefit under 35 U.S.C. .sctn.119(a) of
European Patent Application No. 09157850.0, filed Apr. 14, 2009,
said patent applications hereby fully incorporated herein by
reference.
FIELD OF THE INVENTION
[0002] The present invention relates to the treatment of samples
with focused acoustic energy. In particular the invention relates
to a device for irradiating a sample with focused acoustic energy
to treat the sample and a method for irradiating a sample with
focused acoustic energy to treat the sample.
BACKGROUND OF THE INVENTION
[0003] In recent years progress in many aspects of sample-in
result-out devices, also known as micro total analysis systems
(microTAS) or lab-on-a-chip, has generated, for a variety of
reasons, an increasing interest in in-vitro-diagnostic (IVD)
applications. For example the integration and miniaturization
results in systems requiring a relative small, acceptable
contamination risk of the sample, high sensitivity and short
turn-around time of the test and lower costs per test. Furthermore
between sample-input and result generation minimal operator
intervention shall be required. Operator interventions can be done
by relatively unskilled operators and moderate demands on operating
environment.
[0004] Known technologies of treating samples with acoustic energy
may not be appropriate for certain applications like molecular
device applications because after completion of the sonic treatment
no distinction may be made between a leaking cartridge having a
liquid sample inside and the liquid being used by the device
itself. That may be a non-acceptable treatment result within these
devices using such a liquid-coupling.
[0005] Furthermore the pretreatment function including complex
operations like e.g. mixing, is processed separately and
independently from other processing functions. This is opposite to
the general trend in this domain of further miniaturization and
integration. Even more seriously, it contradicts with for example
hospital or lab requirements to have real small size systems,
because of the very limited space available in these settings.
[0006] In addition to that molecular diagnostic tests often include
technologies with complicated piezo arrays, complicated control
systems and complicated electric drivers. These technologies are
expensive, require a lot of technical support and also need much
space.
SUMMARY OF THE INVENTION
[0007] It may be an object of the invention to provide for an
improved treatment of samples.
[0008] Definitions and abbreviations: It shall be noted that in the
context of this invention the following definitions and
abbreviations will be used:
[0009] Dry coupling: The term "dry coupling" will be used in the
context of the invention as a complete transmission of the acoustic
energy through only non-liquid matter from the source to the
sample.
[0010] Acoustic energy: The term "acoustic energy" is in the
context of the invention used as comprising such terms as sonic
energy, acoustic waves, acoustic pulses, ultrasonic energy,
ultrasonic waves, ultrasound, Shockwaves, sound energy, sound
waves, sonic pulses, pulses, waves or any other grammatical form of
these terms.
[0011] Focal region and focal point: "Focal region" or "focal
point" as used in the context of the present invention means that a
region where the acoustic energy converges and/or hits a target or
sample, although that region has not necessarily to be a single
focused point.
[0012] Device: The expression "device" in the context of the
invention includes molecular diagnostic devices as well as other
devices. Applications of the device may e.g. be in healthcare/life
science, food industry, veterinary practice and forensic
applications.
[0013] Sample: It shall explicitly be noted that the term "sample"
may contain samples for molecular analysis being treated with the
device according to the present invention. For example blood,
cultured blood, urine, aspirate, samples with water like viscosity,
heterogeneous samples or samples on a carrier like BAL, sputum,
tracheal aspirate, CSF, swab and/or brush with pathogen.
Nevertheless this does not mean that any other kind of matter,
solid, liquid, gaseous or any combination thereof is excluded from
being a sample and being irradiated with focused acoustic energy by
the invention.
[0014] NA: "NA" will be used for any nucleic acid.
[0015] Source: In the context of the invention the term "source"
will be used synonymously to the term transducer. Additionally any
other apparatus that is able to emit acoustic energy as defined
within the context of the invention is comprised in the source.
[0016] Propagation path: The expression "propagation path"
describes in the context of the invention the way of the acoustic
energy from the source through any combination of at least the
coupler and the cartridge to the sample. Other elements like
lenses, additional couplers may be in the propagation path. Thus in
the propagation path also the intermediate contact layers of these
different elements are passed by the acoustic energy. Additionally
other layers like e.g. the acoustic window or the interface medium
may be comprised.
[0017] Attenuation: The term "attenuation" in the context of the
invention relates to a decrease of the intensity of the generated
acoustic energy. This may be e.g. due to reflection, absorption,
diffraction, or any combination thereof.
[0018] Treatment of the sample: The term "treatment" or "treating"
is used in the context of the invention to describe the interaction
of the focused acoustic energy with the sample. By means of
focusing the acoustic energy onto the sample in various specific
ways sonochemical and/or sonophysical reactions are caused in the
sample to generate functionalities like e.g. mixing, dispersing,
stirring, elution from swabs or brushes, liquefaction, lysing or
cell release. Thereby this definition of "treatment" also describes
the sonophysical and/or the sonochemical interactions during the
process called "pretreatment." In other words "treatment" comprises
amongst other functionalities the "pretreatment" of a sample.
[0019] Process chamber: The expression "process chamber" will be
similarly used to "chamber" and "chamber of the cartridge."
[0020] Ultrasound: The terms "ultrasound" and "ultrasonic" will be
used for cyclic sound pressure with a frequency between 20 kHz and
100 kHz.
[0021] High Intensity Focused Ultrasound (HiFu): The term "HiFu"
will be used in the context of the invention as focused acoustic
field with source frequencies in the range of 0.2 MHz to 10 Mhz,
with amplitudes chosen to be sufficient efficient to create high
pressure shock-waves and/or cavitation in the focal zone. Focal
zone dimensions (length and diameter) are dependent on the source
transducer type (e.g. natural focusing by flat or enforced focusing
by conical/spherical source transducers). Exemplary length-scales
for the indicated frequency range are (sub) millimeters.
[0022] Sample-in result out-system: A system which accepts a (e.g.
biological) sample, does all the required preparation steps to
prepare for detecting any kind of facts, runs the detection and
delivers the detection results. For example a device for molecular
analysis of samples like e.g. blood or other cells can be provided,
that provides for all necessary analysis steps from the supply of
the natural, untreated sample to the result of the analysis.
[0023] Lens: In the context of the present invention the term
"lens" may be used as a component or a system that is enabled to
spread or converge acoustic energy. Any matter being able to
influence the propagation characteristics of the generated acoustic
energy shall be included within the term "lens."
[0024] Interface/Interface medium: In the context of the invention
the propagation path of the acoustic energy may comprise several
components like the source, the full solid coupler and the
cartridge. In order to describe the transitions or areas where
these different elements of the propagation path get in physical
contact to each other the terms interface and interface medium are
used. For example if a coupler is physically contacted with the
cartridge, the interface medium of the coupler describes the
material used in the coupler within this area of the coupler
brought in contact with the cartridge.
[0025] Coupler: The term coupler will be used in the context of the
invention as an element that is part of the propagation path of the
acoustic energy and transfers may be with other elements the
acoustic energy from the source to the cartridge. Furthermore the
term coupler will be used similarly to the term full solid
coupler.
[0026] Solid gel: In the context of the present invention "solid
gel" comprises a gel-forming material only. It is fully solid and
it is at the same time a gel. Liquid substances are fully avoided
within a solid gel. Thus water or hydrogel is avoided when using a
solid gel. Thus the term "gel" is similarly used in the context of
the invention to the term "solid gel."
[0027] It should be noted that embodiments described in the
following similarly pertain to the device for irradiating a sample
with focused acoustic energy and the method for irradiating a
sample with focused acoustic energy. Synergetic effects may arise
from different combinations of the embodiments although they might
not be described explicitly or in detail.
[0028] Further on, it shall be noted that all embodiments of the
present invention concerning a method, may be carried out with the
order of the steps as described, nevertheless this has not to be
the only and essential order of the steps of the method. All
different orders and combinations of the method steps are herewith
disclosed.
[0029] According to a first aspect of the present invention, there
is provided a full solid coupler for a complete dry coupling of
acoustic energy between a source and a cartridge. Accordingly, in a
first exemplary embodiment of the invention a device for
irradiating a sample with focused acoustic energy to treat the
sample is presented, wherein the device comprises an instrument, a
cartridge, a full solid coupler and a source for generating the
acoustic energy. Furthermore the cartridge has a chamber for
receiving the sample and the full solid coupler provides a complete
dry coupling of the acoustic energy between the source and the
cartridge. The instrument and the cartridge are adapted for
inserting the cartridge into the instrument wherein the cartridge
and the instrument are separable.
[0030] In the following possible further features and advantages of
the device according to the first exemplary embodiment will be
explained in detail.
[0031] In other words, by inserting the cartridge into the
instrument a complete dry propagation path for the focused acoustic
energy from the source to the sample is generated. AU different dry
components of the instrument, the cartridge, the full solid coupler
and the source are thus connected in a complete dry manner after
inserting the cartridge into the instrument. The coupler in general
transmits the acoustic energy from one of its end to another. It
shall explicitly be noted, that the full solid coupler is arranged
at the device in such a way, that it complements or completes the
propagation path of the acoustic energy between the source and the
cartridge in a dry way. In other words the propagation path
comprises before insertion of the full solid coupler a first dry
partial propagation path and a third partial propagation path. By
inserting the coupler between these two parts, the missing second
partial path is supplied. The complete propagation path may for
example be formed firstly out of a material attached to a focusing
transducer, secondly out of polymer based coupler and thirdly out
of a foil between the coupler and the cartridge. Thus a complete
dry coupling between the source and the cartridge is achieved. Thus
the full solid coupler does not have to form the whole propagation
path by itself, but if it is desired, an exemplary embodiment of
the invention may realize this.
[0032] Therefore, the use of water or hydrogel or any gel
containing liquid substances is avoided. Thus after a completion of
an irradiation of the sample with the acoustic energy a clear
distinction between a possibly leaking cartridge containing liquid
matter and the coupling media can be made. In other words,
situations with a high contamination risk due to the leakage of a
cartridge may be recognized by a user of the device more clearly
and even faster.
[0033] As the instrument and the cartridge are totally different
components that are physically separated or at least separable the
volume of the sample to be treated can be chosen by selecting
different cartridges. Furthermore the chamber of the cartridge may
not be totally filled with the sample and thus having an additional
air layer within the chamber above the sample. This may arise in
several technical advantages compared to so-called flow through
systems. An exemplary advantage of an air layer above the sample is
that with HiFu vigorous mixing could be introduced, allowing
treatment of sample volumes much larger than the focal zone volume.
For example by creating a fountain of the sample liquid by means of
HiFu irradiation a mixing mechanism via circulation of the sample
liquid that is imminent in the fountain cycle can be supplied.
Thereby the focal zone in which the HiFu energy creates the
fountain may be quite small compared to the sample volume, but
nevertheless a mixing process is initiated by HiFu via the
fountain. Thus the need to irradiate the whole sample volume that
is to be mixed may be avoided by this exemplary embodiment of the
invention. In other words a large sample may be treated by a
relatively small device.
[0034] Additionally HiFu could create a fountain, which may be used
to create cavitation at relative low (reduced) powers. The
cavitation nuclei may be introduced in the sample by the fountain
droplets returning to the liquid which may reduce the
power-threshold compared with homogenous cavity in water with an
order of magnitude. In other words by creating a fountain out of
the sample (e.g. when the sample is a liquid) the minimum power for
the transducer and thus the minimum acoustic energy to be emitted
from the source can be reduced. This may lead to advantages
described in the context of the present invention.
[0035] In other words the fountain could in addition to cause a
mixing in the sample and a reduction of cavitation power threshold
be used for cooling the sample, as the fountain creates much larger
contact surface of the sample with the surrounding air within the
cartridge.
[0036] The physical separation of the cartridge and the instrument
may lead to a non-integrated system which means that the source,
the coupler and the cartridge may be chosen and applied for a
measurement independently from each other. In other words when the
interface between the three constituting parts of the system
(source, coupler and cartridge) is defined an independent choice of
those three constituting parts may be made as long as the choice
fits with the interface.
[0037] Because of the fact, that the size of the cartridge and the
chamber are independent from the size and shape of the source and
of the coupler an enlargement of the cartridge volume is possible
without having the need to change the acoustic characteristics of
the device. A disadvantage of flow through systems compared to this
embodiment of the invention may be that an enlargement of the
chamber is possible without having to increase also the
transducer.
[0038] Additionally it may be relied on focusing onto a focal zone
and avoiding a dependency of an interaction of the acoustic energy
with a wall of the chamber. In other words the walls of the chamber
are not used as a transducer. In contrast to that known systems
have to take into account that resonance frequencies of chamber
walls are functions of geometry and material properties. These
systems have to match that with the source frequency. As it is not
relied on interactions of the acoustic field and the walls in such
a described way, an enlargement of the chamber may be done
independently from the transducer choice.
[0039] As the cartridge is physically separable from the instrument
the cartridge may be a disposable, consumable and removable
cartridge which may lead to a cheap system for analyzing the sample
with focused acoustic energy. After a treatment of the sample the
cartridge may be discarded without having the need to discard the
source or the coupler. Thus a plurality of measurements provided by
one single instrument and one solid coupler and one source for a
variety of different cartridges with different samples is a
possible way using dry coupling.
[0040] The device may further comprise a lens for focusing the
generated acoustic energy onto the sample.
[0041] Furthermore the irradiation of the sample by focused
acoustic energy causes a treatment of the sample.
[0042] The source or transducer could be a flat or curved piezo
transducer operating between kHz up to MHz frequencies. The
diameter of the transducer may be for example between 5 mm and 35
millimeters (mm) to fit with the volume range, for example between
0.2 milliliters (mL) and 10 mL, one would like to process in the
cartridge. The focal length of the transducers may vary from 5 mm
to 80 mm. Transducer electric input power may vary from 2 watts (W)
to 100 W. According to this exemplary embodiment of the invention
the treatment of samples is possible with lower powers compared to
related known technology. Thus heating due to acoustic energy
absorption of circumjacent matter, especially of the matter between
the source and the sample is avoided, enabling the introduction of
dry coupling.
[0043] The transducer may operate in a continuous mode or in a
burst mode. Applied signal to transducer could have different and
varying forms: e.g. sinusoidal, block, triangular, or any
combination thereof. Frequency may be additionally adjusted to
compensate for frequency shift to heating or to switch focal
length.
[0044] The cartridge may have one of the following characteristics:
disposable, consumable, removable, may contain one chamber or a lot
of chambers, may contain one sample or a lot of samples, industrial
applicable. The cartridge material is not limited to but may
further for example be polyethylene (PE), polypropylene (PP),
polyethylene terephthalate (PET); polymethylpentene (PMP),
Polymethylmethacrylate (PMMA), polycarbonate (PC) and polystyrene
(PS).
[0045] In addition to that the cartridge is also a physically
independent device from the coupler. Thus the cartridge is
different and separable from the instrument and also from the
coupler. This exemplary embodiment of the invention does not
exclude, that the coupler is placed or fixed onto the cartridge or
the instrument, but contains this possibility.
[0046] One main advantage is that all desired and needed processing
of the sample may be done in one single chamber of the cartridge.
Furthermore the whole processing by the applied acoustic energy may
be done according to the sample-in result-out principle with all
necessary actuation coming from one single source of the device. By
means of the focused acoustic energy the sample may be treated with
a lot of different functionalities like sample pretreatment and
lysis in one single chamber being a process chamber. Especially
HiFu may be used for these processes.
[0047] To achieve a high intensity of the acoustic energy at the
receiving position (chamber in cartridge and thus at the sample) it
is preferred that the focus quality of the source or transducer
and/or lens is sufficient, that the acoustic attenuation of the
materials in the propagation path of the acoustic energy is
sufficiently low which means a low impedance and/or a low
thickness, and that reflection at the material interfaces in the
propagation path of the acoustic energy is sufficiently low which
means for the dry coupler that the thickness and roughness of the
two contacting layers should be sufficiently small. This exemplary
embodiment of the invention meets these requirements.
[0048] Power may be supplied to the source from the instrument via
e.g. leads or brushes. The full solid coupler may comprise
different pieces, parts or segments.
[0049] Furthermore the dry coupling may induce, that on the
microscale the contact between for example the source and the
coupler (first layer) and/or the coupler and the cartridge (second
layer) may approach direct contact condition, in other words as
close as possible in order to achieve efficient dry coupling. Thus
the surfaces of the two layers may be on the microscale or
nanoscale as conformal as possible to minimize or eliminate
air-pockets between the two layers in dry contact.
[0050] In other words to minimize or eliminate air pockets the
following requirements may be met by the device: The surface
roughness may be sufficiently low of the source, the coupler, the
cartridge, the full solid coupler and an interface medium. Also the
used materials may be sufficiently "flexible" to achieve
conformality. Thereby a conformality order may be considered
liquids>hydro gels>solid gels>rubbers>(elastic)
foils>thermoplastic polymer>thermo harders, metals, ceramics
and other solid materials.
[0051] The acoustic energy or acoustic radiation may propagate
through a first part of the path unfocused and may later be focused
within a second part of the path to propagate focused through a
third part of the path till the sample. Previous or subsequent
focusing is also possible.
[0052] The required power for creating a cavitation process in the
sample may be reduced by this exemplary embodiment of the
invention, as additional nucleation sites may be introduced in the
chamber (e.g. an element with an appropriate high surface roughness
e.g. a rod) or a fountain may be induced. Droplets falling back
from the fountain into the sample may reduce this power threshold.
As the present construction enables both possibilities low power
HiFu may be used for preparing and treating the sample.
[0053] As the required power may be decreased by the present
invention, additional refraction, being generated at high
intensities, may be avoided.
[0054] According to another exemplary embodiment of the invention
the focused acoustic energy is high intensity focused ultrasound
(HiFu).
[0055] Thereby the source frequencies may be in the range of 0.2
MHz to 10 MHz, with amplitudes chosen to be sufficient efficient to
create high pressure shock-waves and/or cavitation in the focal
zone. Focal zone dimensions may be dependent on the source
transducer type. Exemplary length-scales for the indicated
frequency range are (sub) millimeters. Furthermore flat or curved
piezo transducer may be used operating between 0.2 MHz and 10 MHz,
or between 0.75 MHz and 3 MHz or between 1 MHz and 2 MHz. The
diameter of the transducer may be for example between 5 mm and 35
mm to fit with the volume range (0.2 mL-10 mL) one would like to
process in the cartridge. The focal length of the transducers may
vary from 5 mm to 80 mm. Transducer electric input power may vary
from 0.5 W to 100 W.
[0056] In other words, this exemplary embodiment of the invention
may be used as a HiFu molecular device for treating and/or
analyzing molecular samples. Thereby no liquid matter must be used
for coupling the acoustic energy from the source to the sample.
Thus liquid contamination risks may be reduced and by using
disposable or consumable cartridges an uncomplicated, cheap and
fast way of measuring characteristics of the sample plus preparing
the sample with the device and thus with HiFu may be provided.
[0057] Due to the relatively short wavelength of HiFu compared to
ultrasound, an enhanced focusing onto a smaller region is possible.
This leads to a miniaturization advantage.
[0058] In addition to that various different focal region shapes
may be used for treating the sample by the HiFu.
[0059] As HiFu enables a user to treat a sample e.g. with
functionalities like mixing with a reagent, circulation, release of
a cell, pathogen and matrix from a swab, release of a cell,
pathogen and matrix from a brush, liquefaction, incubation of the
sample with a reagent at room temperature or elevated temperature,
shaking, mixing; stirring, extraction, NA extraction, flow
generation, sample homogenation, separating by centrifuging, and
any combination thereof, lysis, lysis of microorganisms, incubation
of the sample with a reagent at room or elevated temperature, and
any combination thereof a huge variety applications for the device
is created.
[0060] Furthermore known systems may be limited dictated by physics
because real miniaturization of the ultrasound transmitter may not
be possible; known systems may thus be limited to about 100 mm.
This embodiment of the invention may be miniaturized smaller than
100 mm.
[0061] Further on another disadvantage of known systems may be that
the resonance frequency of the ultrasound chamber is design and
material dependent and should be matched with the chosen ultrasound
transmitter frequency. Manufacturing tolerances may have to include
this dependency. In contrary to that, any resonance frequency of
the device may not have been taken into account, as described
above.
[0062] Furthermore other instruments using acoustic energy may be
limited to a small volume chamber as according to basic physical
laws of mechanics an increasing in dimensions means a reduction of
the resonance frequency of the chamber or the system. Parallel
existing requirements of the ultrasound frequency originating in
the specifications of the sample may thus make it useless to
increase the size of the chamber. This may limit the spectra of the
applications of such a known instrument.
[0063] In contrary to that, a non integrated system is presented,
in which the cartridge is physically independent i.e. separated
from the source and the coupler, as described above. It may be that
no resonance frequency of the chamber has to be taken into account,
when selecting the desired size of the cartridge or the chamber.
This is an important advantage above the known technology.
[0064] Furthermore this exemplary embodiment is enabled to avoid,
if necessary, a flow-through technique, which may complicate the
combination with incubation at elevated temperature. In addition
these flow-through technologies may have the need to provide some
kind of beads to the chamber. But in the case a flow through may be
desired, the present idea is able to provide for that.
[0065] In other words this exemplary embodiment of the invention
distinguishes from technologies using ultrasound impacting the wall
of the chamber. In these known systems the resonance frequency is
dependent from geometry and/or the material of the device.
[0066] Furthermore in contrast to flow-through systems which use
homogenous cavitation, the power can be reduced in this exemplary
embodiment, as this exemplary embodiment of the invention may
supply for an air layer in the chamber, which makes it possible to
introduce nucleation sites or to produce a fountain as described
above. By means of additional nucleation sites like a rod that is
introduced in the chamber or by means of the described fountain,
the power threshold to initiate cavitation may be reduced.
Furthermore it may be provided for an incubation possibility of the
sample although not all of the sample fluid has to be in the focal
zone.
[0067] This may enable the user to use smaller transducers and less
power which enables the introduction of the full solid coupler or
dry coupling. Furthermore the combination of incubation may be
facilitated.
[0068] In addition to that this exemplary embodiment of the
invention is able to use additional different functions, e.g.
elution of swabs in a process chamber. As HiFu is used with dry
coupling it allows to detect cartridge leakage and therefore
contamination could be detected in an early stage.
[0069] According to yet another embodiment of the present invention
a source is one of part of the instrument or part of the
cartridge.
[0070] In a first example of this embodiment the source may be
implemented in the instrument of the device. Thus a plurality of
cartridges may be irradiated one after another by one and the same
acoustic energy source. Thus measurement results of different
cartridges may be more comparable and reliable as deviations
originating from different sources can be excluded.
[0071] In a second example of this embodiment of the invention the
source is part of the cartridge. For example, a cartridge may be
provided with a source and a full solid coupler being situated
between the source and the cartridge. For example, they may be
glued together to one unit. Also other fixation possibilities shall
be comprised. By inserting this unit into the instrument the
electric connection between the power supply for the source is
plugged together. Thus a complete dry coupling is generated in this
embodiment of the invention.
[0072] By means of the integration of the source into the cartridge
a pre-selection or pre-adaption of the specific source for certain
measurement intentions is possible. Thus in combination with the
instrument different types of cartridges with specifically selected
sources for these cartridges and for specific measurements may be
used with one single instrument. This means an increase operation
field of the instrument. In addition to that the cartridges and the
sources being attached to the cartridges may be disposable and thus
may provide a cheap and uncomplicated solution for treating
different samples in different cartridges with different sources
attached by one single instrument.
[0073] According to another exemplary embodiment of the invention,
the instrument and the cartridge are arranged in combination in
such a way that by inserting the cartridge into the instrument the
propagation path for transmitting the acoustic energy from the
source to the sample is formed wherein the propagation path
comprises only of non-fluidic matter.
[0074] In other words, the interaction of the cartridge and the
instrument during the insertion process yields to the complete dry
coupling propagation path. Therefore, the corresponding surfaces of
the instrument and the cartridge are brought together during an
insertion process and they may be shaped in a for example
form-closed way or in a force-fit way. In addition to this shape
fitting of the contour of the instrument and the cartridge extra
means for applying a pressure between these elements and the
additional coupler may be provided. In other words, only solid
materials or gaseous materials, like air pockets, are present in
the propagation path of the acoustic energy.
[0075] According to another exemplary embodiment of the invention,
the full solid coupler is formed out of the material selected from
the group comprising solid gel, rubber, elastic foil, polymer based
material, thermoplastic polymers, polymer having a low acoustic
attenuation characteristic, metal, semiconductor, ceramic,
polypropylene, aluminum, and a stack of these materials.
[0076] It shall explicitly be noted, that the full solid coupler
may be formed out of a polymer based material.
[0077] The used materials may obey elastic characteristics that
allow a conformable adaption of the coupler to the shape of a
component of the device for example of the cartridge or of the
source. Thereby the material of the full solid coupler may be
chosen in such a way, that air pockets at any interface within the
propagation path are minimized or avoided to achieve efficient dry
coupling. Furthermore the full solid coupler may also contain the
above mentioned materials as partial components and other not
mentioned materials may be contained in the full solid coupler.
[0078] Calculations have shown that the stack could increase the
amount of energy which could be transferred to the receiving
position, but at the expense of a more complicated coupler. In
other words impedance matching may be used. Thus the full solid
coupler may comprise several components, that together yield to a
complete and efficient dry coupling of the acoustic energy from the
source to the sample.
[0079] According to another exemplary embodiment of the invention,
the cartridge comprises an acoustic window wherein the acoustic
window is made of a flexible foil and wherein the full solid
coupler is physically contacted with the acoustic window by
inserting the cartridge into the instrument.
[0080] In order to achieve a high intensity of the focused acoustic
energy at the receiving position (chamber in the cartridge where
the sample is positioned) it may be essential that the attenuation
of the materials in the transport path of the HiFu is sufficiently
low. Furthermore air pockets shall be minimized or eliminated by
using surface roughness's that are sufficiently low. Also materials
that are sufficiently flexible to achieve conformity may be used.
These requirements may be met by the acoustic window that is made
out of a flexible material like a plastic foil. Thereby the plastic
foil may adapt its shape during an insertion of the cartridge into
the instrument to the shape of the contact surface of the cartridge
or the shape of the full solid coupler.
[0081] The acoustic window of the cartridge may be sufficiently
large that the cross section of the HiFu cone at the chosen
acoustic window distance fits completely in the window. The
acoustic window could be flat or curved. The acoustic window is of
made of a thin layer of a low attenuation polymer, e.g. PP, PMP. It
is also important that the remaining part of walls of the lysis
chamber below the fluid level may be sufficiently thin to reduce
acoustic losses and to limit heating of the chamber housing
[0082] According to another exemplary embodiment a contact pressure
between the full solid coupler and the cartridge is applied,
wherein the contact pressure is generated by at least one method
from the group comprising applying over pressure in the chamber of
the cartridge, applying local under pressure outside of the
cartridge, and pressing the cartridge and the full solid coupler
against each other by means of a force.
[0083] The contact pressure between the solid coupler and the
cartridge surface is applied in a sufficient way to get rid of air
or air pockets at the interface or at any intermediate layer in the
propagation path of the acoustic energy. Pressing e.g. a convex
shaped solid coupler against a flat cartridge, with the cartridge
material being sufficiently flexible to become conformal to the
shape of the solid coupler may be possible solution. In addition to
that the coupler may also have such a flexibility.
[0084] Another exemplary embodiment may be a dry interface solution
comprising a smooth spherical or conical shaped HiFu transducer and
a flexible cartridge foil.
[0085] Thereby the contact pressure yields to a force that presses
a least the three components source, coupler and cartridge together
in such a way, that air pockets may be minimized between some or
all intermediate contacting surfaces. Therefore especially smooth
and flexible materials may be used for these surfaces.
[0086] According to another exemplary embodiment of the invention,
the full solid coupler has a first contact surface for contacting
the acoustic window and the cartridge has a second contact surface
for contacting the acoustic window. Furthermore at least one of the
first contact surface, the second contact surface and the acoustic
window has a surface roughness value selected from the group
comprising smaller than 0.5 micrometers (.mu.m), smaller than 1
.mu.m, and smaller than 2 .mu.m.
[0087] Due to this embodiment of the invention air pockets and thus
transmission losses in the propagating acoustic energy may be
minimized or eliminated.
[0088] An interface medium between the instrument and cartridge to
enable acoustic energy transport across dry interface may be made
of a low attenuation material like rubber (e.g. RT 615), (elastic)
foil (e.g. PP, PP based Thermoplastic Elastomer, PMP), or
thermoplastic polymer (e.g. PP). The interface layer could be part
of the instrument or of the cartridge. For example the cartridge
bottom layer contacting the coupler could also be at the same time
the interface medium.
[0089] According to another exemplary embodiment of the invention,
the propagation path has a gradient of acoustic impedance, wherein
the gradient is monotonously decreasing in a direction from the
source to the sample.
[0090] This embodiment may lead to further reduction of acoustic
energy losses, as a coupling from one component to another
component in the propagation path may be improved due to the
gradient of the acoustic impedance. By applying such an acoustic
impedance profile within the propagation path reflection and
absorption of the focused acoustic energy may be reduced. This may
lead to a better yield or spoil of a given power.
[0091] The acoustic impedance of the materials used within the
propagation path of the acoustic energy is going from relative high
on the side of the source to relative low at the sample/cartridge
site. In addition to that principle laws of acoustics may be used
to optimize the choices of dimensions and material of the device
and its components.
[0092] According to another exemplary embodiment of the invention,
the full solid coupler is selected from the group comprising a
coupler being a physically separate component placed between the
source and the cartridge, a coupler being part of the source, a
coupler being part of the cartridge and any combination
thereof.
[0093] For example, a configuration with the source being a piezo
transducer combined with a metal lens on top that has a polymer
coupler on top of the metal lens is possible. Also a curved source
working simultaneously as a lens may be provided with a polymer
coupler on top of that curved source. The coupler may physically be
bonded to the source or the cartridge but it can also be hold on
top of one of these components by external pressure applied to
these components. Referring now to the following FIGS. 10 to 14 a
large variety of combinations of arranging and fixing the coupler
between the source and the cartridge are possible. Placing the
coupler on the source, on the cartridge, on a lens, on a second
additional coupler and on an acoustic window with different
fixation possibilities like pressing together, gluing together,
depositing a coupler on a component, and any combination thereof
are comprised within this embodiment of the invention.
[0094] According to another exemplary embodiment of the invention,
a lens for focusing the generated acoustic energy onto the sample
is further comprised. Thereby the lens is selected from the group
comprising a lens being a physically separate component placed
between the source and the cartridge, a lens being part of the
source, a source with a focusing shape being the lens, an array of
sources that yield to a focus acoustic energy, a lens being part of
the cartridge, a lens made out of a polymer, having a low acoustic
attenuation characteristic, a metal lens, a ceramic lens, a
polypropylene lens, an aluminum lens, a hybrid lens and any
combination thereof.
[0095] Lens may be made of a low attenuation polymer, metal or
ceramic. For environmental reasons the lens may be integrated in
the consumable and made of a polymer, e.g. PP.
[0096] As a first characteristic of the lens the lens is able to
focus the generated acoustic energy onto the sample. In order to
reduce the transmission losses the lens may be attached to the
source. For example a metal lens may be fixed onto a piezo
transducer yielding to the emission of a focused acoustic field.
Furthermore an array of a plurality of sources may be spatially
placed in such a way and electronically driven in such a way that
the superposition of all the singular acoustic fields yields a
focused acoustic field. Furthermore it is possible that the lens is
part of the cartridge for example being fixed to the bottom of the
cartridge. In addition to that this example may further comprise a
source being part of the cartridge.
[0097] In order to create multi-focality also a hybrid lens may be
used in this exemplary embodiment of the invention. Thereby the
lens has at least two different emitting zones which means, that
the different emitting zones of the lens deviate from each other by
at least one of the following components shape, surface roughness,
material, and any combination thereof. To shortly summarize the
function of a hybrid lens it has to be said that an incoming
homogeneous acoustic field will be transferred by the hybrid lens
into a non-homogenous acoustic field having for example two
different focal regions.
[0098] According to another exemplary embodiment of the invention,
in the one single chamber of the cartridge pretreatment and lysis
are applied to the sample by means of the focused acoustic energy.
Thereby pretreatment is a method selected from the group comprising
mixing with a reagent, circulation, release of a cell, pathogen and
matrix from a swap, release of a cell, pathogen and matrix from a
brush, liquefaction, incubation of the sample with a reagent and/or
enzyme at room temperature or elevated temperature, shaking,
mixing, stifling, extraction, NA extraction, flow generation,
sample homogenation, separating by centrifuging and any combination
thereof. Furthermore lysis is a method selected from the group
comprising mixing with a reagent, mixing with a reagent different
to the reagent applied during pretreatment, circulation, lysis of
microorganisms, incubation of the sample with a reagent at room or
elevated temperature or a temperature different from the
temperature applied during pretreatment and any combination
thereof.
[0099] It shall explicitly be noted that this combination of
pretreatment and lysis in one single chamber by means of the
focused acoustic energy originating from only one single source may
be applied without providing a dry coupling. No full solid coupler
or a propagation path out of completely dry media is necessary.
[0100] Accordingly, a second aspect of the present invention is
directed to the application of pretreatment and lysis to the sample
by means of focused acoustic energy in the one single chamber of
the cartridge, i.e. in particular in the same chamber. An exemplary
embodiment of this aspect of the present invention provides a
device for irradiating a sample with focused acoustic energy to
treat the sample, the device comprising an instrument, a cartridge,
and a source for generating the acoustic energy. The cartridge has
a chamber for receiving the sample. The instrument and the
cartridge are adapted for inserting the cartridge into the
instrument. The cartridge and the instrument are separable. The
device is designed such that pretreatment and lysis are applicable
to the sample in the chamber of the cartridge by means of the
focused acoustic energy.
[0101] In addition to that an exemplary embodiment of this second
aspect of the present invention further relates to a corresponding
instrument for irradiating a sample with focused acoustic energy to
treat the sample, the instrument comprising a source for generating
the acoustic energy. The instrument is adapted to receive a
cartridge being separable from the instrument, the cartridge
providing a chamber for receiving the sample. The instrument is
designed such that, when the cartridge is being inserted in the
instrument, pretreatment and lysis are applicable to the sample in
the chamber of the cartridge by means of the focused acoustic
energy.
[0102] Correspondingly, a cartridge is provided in another
exemplary embodiment which cartridge for an instrument for
irradiating a sample with focused acoustic energy generated by a
source to treat the sample comprises a chamber for receiving the
sample. The cartridge is adapted for being inserted into an
instrument and being separable from the instrument. The cartridge
is designed such that when being inserted into the instrument
pretreatment and lysis are applicable to the sample in the chamber
by means of the focused acoustic energy.
[0103] In addition to that exemplary embodiments of this aspect of
the invention further relate to a corresponding method for
pre-treating and lysing a sample in one single chamber by means of
focused acoustic energy like for example HiFu originating from one
single source, preferably by such device, and a computer program
element characterized by being adapted when being used to control a
device for pre-treating and lysing a sample to cause the device for
performing the steps of this corresponding method.
[0104] These exemplary embodiments may for example combine
pretreatment and lysis in a single chamber by using single focus
HiFu, but also the use of multi focus HiFu is possible. But also
any combination with incubation is possible.
[0105] In other words, manual steps for doing sample pretreatment
can be avoided by an exemplary embodiment of the device, the
corresponding instrument and cartridge, the corresponding method
and the computer program element. Pretreatment is integrated in the
cartridge to increase ease-of-use and to decrease fluidic
interfacing with the external world and contamination risk.
Furthermore pretreatment and lysis functions are integrated in one
single chamber that could be exposed to HiFu and/or heated and/or
cooled to reduce complexity, costs and size of the device and the
procedure for doing treatment and lysis together. Pretreatment and
lysis functions advantageously are processed without the sample
leaving the chamber in between, and/or advantageously are processed
in a fully automated manner, and/or advantageously are processed
sequentially or simultaneously.
[0106] This second aspect of the invention may be used for any
application requiring pretreatment and/or lysis. Applications may
not be limited to healthcare, life science, food industries and
veterinary practice. This relates to any embodiment of the
invention.
[0107] Especially for lysing difficult micro-organisms the state of
the art technique of applying thermal lysis has several
insufficiencies. In contrary to that this aspect of the present
invention uses focused acoustic energy, especially HiFu for solving
these problems. By means of such a fully integrated in vitro
preparation and detection instrument a system for sample-in
result-out tests is provided, especially for nucleic acid (NA),
protein or cell detection. Furthermore nucleic acid analysis,
protein analysis and cell analysis may be possible by a so called
micro total analysis system.
[0108] Additionally, existing lysing methods comprise grinding or
bead beating which may be avoided here.
[0109] In general nucleic acid sample preparation protocols are
more complicated than cell or protein preparation protocols.
Although this aspects of the invention may be for the major part on
nucleic acid sample preparation it is not limited to this.
[0110] For this reason a single solution with a high flexibility is
needed to accommodate these deviations in needed pretreatment. This
aspect of the invention meets these requirements with high degree
of flexibility on pretreatment and lysis protocols.
[0111] It shall be noted that preferred embodiments of other
aspects of the present invention shall be considered as preferred
and disclosed embodiments with respect to the present aspect, too,
and vice versa.
[0112] According to another exemplary embodiment of the invention,
the device is adapted in such a way that it generates at least two
different focal regions at the sample.
[0113] It shall explicitly be noted that this exemplary embodiment
of the invention may be applied or implemented without having the
need to provide for the complete dry coupling features. In other
words, the creation of a multi-focality by the device may also be
used in combination with non-solid coupling matter.
[0114] Accordingly, a third aspect of the present invention is
directed to the generation of two different focal regions at the
sample. In an exemplary embodiment of this third aspect of the
present invention a device is presented for irradiating a sample
with focused acoustic energy to treat the sample comprising an
instrument, a cartridge, and a source for generating the acoustic
energy. The cartridge has a chamber for receiving the sample. The
instrument and the cartridge are adapted for inserting the
cartridge into the instrument. The cartridge and the instrument are
separable. The device is designed for generating at least two
different focal regions of acoustic energy at the sample.
[0115] In addition to that an exemplary embodiment of the third
aspect of the present invention further relates to a corresponding
instrument for irradiating a sample with focused acoustic energy to
treat the sample, the instrument comprising a source for generating
the acoustic energy. The instrument is adapted to receive a
cartridge being separable from the instrument and providing a
chamber for receiving the sample. The instrument is designed for
generating at least two different focal regions of acoustic energy
at the sample when the cartridge is inserted in the instrument.
[0116] Correspondingly, a cartridge is provided in another
exemplary embodiment which cartridge for an instrument for
irradiating a sample with focused acoustic energy generated by a
source to treat the sample comprises a chamber for receiving the
sample. The cartridge is adapted for being inserted into an
instrument and being separable from the instrument. The cartridge
is designed for allowing generating at least two different focal
regions of acoustic energy at the sample when being inserted in the
instrument.
[0117] Furthermore it shall explicitly be noted that a
corresponding method for generating at least two different focal
regions at the sample by the device and a corresponding computer
program element for controlling a device generating a
multi-focality to the sample is comprised within this embodiment.
Thereby the computer program element may be characterized by being
adapted when in use on a device for creating a multi-focality to
the sample to cause the device for performing the steps of the
corresponding method.
[0118] In other words, a treatment protocol using two different
focal zones for providing different focus conditions is provided.
For example, focus conditions for doing mixing a liquid circulation
by means of focused acoustic energy may be different from the
requirements for doing lysis with for example microorganisms. This
embodiment of the invention meets these requirements.
[0119] By providing at least two different focal regions at the
sample the device may provide for attractive simple and cheap
molecular diagnostic tests. Furthermore complex arrangements of
piezo arrays, complicated systems and/or drivers can be avoided by
this exemplary embodiment of the invention. Furthermore the
integration of several different functionalities (like e.g. mixing,
circulating, and lysing) into one chamber that are processed by the
two different focal regions a miniaturization of the molecular
diagnostic device is possible.
[0120] In other words the molecular diagnostic device is a
multi-focality HiFu molecular diagnostic device for applying
different focal regions to the sample. This can be used for
generating and combining different treatment functionalities. For
example point-like focused HiFu may be optimal for doing lysis and
zone-like focused HiFu may be optimal for mixing and/or
circulating. Thereby point like means a comparatively small focal
region, and a zone-like focus means a comparatively large focal
region. Different focal regions may also differ in shape and in
size. These different focal requirements are met by this exemplary
embodiment of the invention.
[0121] Furthermore lysis by means of HiFu requires high acoustic
pressures. High pressures are achieved by good quality focusing
which is achieved by this exemplary embodiment of the invention by
means of a first highly focused part of the generated acoustic
energy. In contrary to that to release particles or cells from
swabs, to release and homogenize feces from a carrier, e.g. swabs,
brush, to homogenize liquid present in the chamber with reagents
added to the chamber mixing and circulation may be required in one
single chamber of the cartridge. Thus a second part of the
generated acoustic energy is focused to a comparatively large,
zone-like second focal region at the sample. Thus two different
treatment functionalities may be applied to the sample during the
same time, in one single chamber and without any manual
intervention of a user.
[0122] According to another exemplary embodiment of the invention,
the at least two different focal regions are generated by means of
an element selected from the group comprising a plurality of
sources, a single source and a hybrid lens, one single source with
different roughness zones and one single source being excited
differently at different positions of the source, and any
combination thereof. Such element may be embodied as an element
external to the cartridge, or an element belonging to the
cartridge, or as an element integrated into the cartridge.
[0123] A plurality of sources comprises at least two single
sources, as well as an array of sources being electronically
controlled in such a way that the superposition field of all the
sources yields to a total field having at least two focal regions.
Furthermore the hybrid lens may consist of a moderately focusing
material and a highly focusing material. These materials may be
positioned at different parts of the lens yielding to
multi-focality. For example a concave shaped hybrid lens may be
attached to a flat source like a transducer. But also a curved
transducer with a curved hybrid lens made out of a moderately
focusing material and a highly focusing material is possible. In
order to find an optimized distribution of these two different
materials acoustic modeling may be performed on different
configurations. For example, the lens may be formed out of
polypropylene. Furthermore lens radius may vary due to the
application of the device. In order to create a multi-focality at
the receiving position where the sample is located, the source may
also be provided with different roughness zones, which means that
the surface of the source obeys different surface roughness
values.
[0124] The different emitting zones, more detailed the respective
surfaces of these zones, may have different roughness properties.
These different roughness properties yield to different acoustic
irradiating characteristics of the zones, which leads to at least
two different focal zones. Thereby the source or transducer itself
may have these zones. But also an additional component may be added
on top of the transducer, wherein the component obeys these
different surface roughness characteristics. In other words the
gist of this possibility is that the surface of the transducer is
segmented in a smooth and rough area delivering respectively highly
and moderately focused acoustic energy, especially HiFu, to the
sample.
[0125] It shall be noted that preferred embodiments of other
aspects of the present invention shall be considered as preferred
and disclosed embodiments with respect to the present aspect, too,
and vice versa.
[0126] According to another exemplary embodiment of the invention,
the focused acoustic energy is used for reducing the viscosity of
the sample.
[0127] It shall explicitly be noted that this embodiment of the
invention does not necessarily need to contain all the dry coupling
features. In particular no full solid coupler or a completely dry
propagation path is necessary.
[0128] Accordingly, a fourth aspect of the present invention is
directed to using the focused energy for reducing the viscosity of
the sample. In an exemplary embodiment of this fourth aspect of the
present invention a device is provided for irradiating a sample
with focused acoustic energy to treat the sample comprising an
instrument, a cartridge, and a source for generating the acoustic
energy. The cartridge has a chamber for receiving the sample. The
instrument and the cartridge are adapted for inserting the
cartridge into the instrument. The cartridge and the instrument are
separable. The device is designed for using the focused acoustic
energy for reducing the viscosity of the sample.
[0129] In addition to that an exemplary embodiment of the fourth
aspect of the present invention further relates to a corresponding
instrument for irradiating a sample with focused acoustic energy to
treat the sample, the instrument comprising a source for generating
the acoustic energy. The instrument is adapted to receive a
cartridge being separable from the instrument and providing a
chamber for receiving the sample. The instrument is designed for
using the focused acoustic energy for reducing the viscosity of the
sample when the cartridge is inserted into the instrument.
[0130] Correspondingly, a cartridge is provided in another
exemplary embodiment which cartridge for an instrument for
irradiating a sample with focused acoustic energy generated by a
source to treat the sample comprises a chamber for receiving the
sample. The cartridge is adapted for being inserted into an
instrument and being separable from the instrument. The cartridge
is designed for allowing reducing the viscosity of the sample by
means of focused acoustic energy applied to the sample when being
inserted into the instrument.
[0131] In addition to that an exemplary embodiment comprises a
corresponding method for reducing the viscosity of the sample,
preferably by such device, and a corresponding computer program
element. Thereby the computer program element is characterized by
being adapted when in use on a device for reducing the viscosity of
the sample by means of irradiating the sample with focusing
acoustic energy to cause the device for performing the steps of the
corresponding method.
[0132] In order to reduce the viscosity of a sample like for
example BAL, sputum, blood, feces, or any other sample present on a
swab this embodiment of the invention suggests to use focused
acoustic energy for example HiFu to cause this reduction. This
method may be implemented in a complete sample-in result-out
solution in which a subsequent pretreatment and lysis of the sample
may be possible in the one chamber of the cartridge. Thus by means
of only one single source a complete process of viscosity
reduction, further pretreatment and lysis is possible.
[0133] For example, a source having the following characteristics
may be used for the reduction of the sample viscosity. 3.0 MHz
transducer with a diameter of 25 mm a focal length of 22 mm.
Furthermore the bottom of the cartridge may be set at 15 mm
distance of the transducer. Exemplary power of 5 W may be applied
to the sample for approximately 300 s. By means of such a HiFu
application the sample may be more homogeneous after such a HiFu
exposure and the viscosity may drop from the original viscosity to
for example a water-like viscosity. Thus it can be concluded that
HiFu forces combine the ability to reduce the molecular weight of
the macromolecules and as a result the viscosity and the ability to
circulate and mix a sample in a process chamber.
[0134] It shall explicitly be noted that this exemplary embodiment
of the invention may be used for any application requiring
circulation and/or mixing in the sub-millimeter volume range in a
device. The applications may be also in the life sciences,
lab-on-the-chip, and mTAS applications.
[0135] It shall be noted that preferred embodiments of other
aspects of the present invention shall be considered as preferred
and disclosed embodiments with respect to the present aspect, too,
and vice versa.
[0136] According to another exemplary embodiment of the invention,
a detection unit for applying measurements on the sample is further
comprised. Thereby the irradiation of the sample with the focused
acoustic energy leads to a treatment of the sample.
[0137] In other words, this exemplary embodiment of the invention
provides for a complete sample-in result-out system, where no
manual step has to be done by the user. A sample may be inserted
into the device and by means of the focused acoustic energy the
sample is treated in a desired way. Subsequently or also previously
measurements may be applied to the sample by means of the detection
unit. Thereby the device is enabled to deliver the measurement
results to for example a user interface to the user. For example,
functionalities like liquefaction, stirring, mixing, circulation,
pretreatment, incubation and lysis may be done before or after any
measurement of the detection unit by means of the focused acoustic
energy. A fully automated system is thus provided to the user.
[0138] It shall further be noted that this exemplary embodiment of
the invention may not necessarily contain all dry coupling
features. In particular, no full solid and dry coupler or a
completely dry propagation path is necessary.
[0139] Accordingly, a fifth aspect of the present invention is
directed to a detection unit for applying measurements on the
sample. In an exemplary embodiment of this fifth aspect of the
present invention a device is provided for irradiating a sample
with focused acoustic energy to treat the sample comprising an
instrument, a cartridge, and a source for generating the acoustic
energy. The cartridge has a chamber for receiving the sample. The
instrument and the cartridge are adapted for inserting the
cartridge into the instrument. The cartridge and the instrument are
separable. The device comprises a detection unit for applying
measurements on the sample.
[0140] In addition to that this exemplary embodiment of the
invention further relates to a corresponding instrument for
irradiating a sample with focused acoustic energy to treat the
sample, the instrument comprising a source for generating the
acoustic energy. The instrument is adapted to receive a cartridge
being separable from the instrument and providing a chamber for
receiving the sample. The instrument comprises a detection unit for
applying measurements on the sample when the cartridge is inserted
into the instrument.
[0141] Correspondingly, a cartridge is claimed in another exemplary
embodiment which cartridge for an instrument for irradiating a
sample with focused acoustic energy generated by a source to treat
the sample comprises a chamber for receiving the sample. The
cartridge is adapted for being inserted into an instrument and
being separable from the instrument. The cartridge is designed such
that when being inserted into the instrument a detection unit may
apply measurements on the sample.
[0142] In addition to that it shall be noted that this exemplary
embodiment comprises a corresponding method for applying
measurements on the sample by such device and a corresponding
computer program element. Thereby the computer program element is
characterized by being adapted when in use on such a sample-in
result-out system to cause the device for performing the steps of
the corresponding method.
[0143] This allows in vitro treatment of the sample by means of
e.g. HiFu and at the same time in vitro detection which leads to a
complete sample-in result-out system.
[0144] Especially for a molecular device being a device according
to an embodiment of the invention and which device is enabled to
extract, purify, amplify and detect nucleic acids it shall be
stated the following: Extraction and/or purification of nucleic
acids is based on adsorption and/or desorption on a solid surface.
Any surface offering sufficient capture area should be regarded as
a part of an embodiment of the invention. Common surface capture
embodiments are (for example magnetic) particles and membranes. Any
capturing material capable of delivering nucleic acids of
sufficient quality for multiplication purposes should be regarded
as part of an embodiment of the invention. Widely used materials
are e.g. silica, magnetized silica, iron-oxide, aminogroup
functionalized polystyrene. Also other materials are possible.
[0145] The detection unit and thus the detection method of choice
may be dependent on the application area like e.g. nucleic acids,
protein or cell detection.
[0146] For nucleic acid amplification and detection e.g. a large
number of isothermal and thermal cycling amplification methods are
described. Polymerase chain reaction (PCR) is one of the most used
methods. The sample-in result-out system according to this
exemplary embodiment of the invention implements such a PCR
functionality into the chamber where the sample is also treated by
means of HiFu.
[0147] PCR is further subdivided in two subcategories namely
end-point and real-time PCR (rtPCR). Of these two rtPCR is most
widely used (rtPCR amplification is running in parallel with
detection). For detection of nucleic acids one may for example use
detectable markers such as fluorescent markers which may be
incorporated in the amplified nucleic acids during PCR. Other
detectable labels or even label-free methods may also be used.
[0148] For protein detection, common approaches such as a
combination of antibody capture and optical readout, e.g.
fluorescence, of magnetic readout may be used.
[0149] For cell detection, optical methods as they are widely used
to count, analyze cell shape, etc, but (di) electrophoretic and
electrical properties could also used to detect/characterize
cells.
[0150] All the before mentioned detection possibilities of this
embodiment of the invention correspond to the detection unit that
is used in this embodiment of the invention. Thus the realized
sample-in result-out system may incorporate any of these detection
or measurement features.
[0151] According to another exemplary embodiment of the invention,
the detection unit is for applying at least one measurement to the
sample selected from the group comprising optical measurements,
magnetic measurements, thermal measurements, electrical
measurements, chemical measurements, sonic measurements, and any
combination thereof.
[0152] The device may further comprise at least one of: an
extraction unit; a nucleic acid amplification unit; a reagent
storage unit; a detection unit a detection unit for applying
measurements on the sample wherein the detection unit is for
applying at least one measurement to the sample selected from the
group comprising optical measurements, magnetic measurements,
thermal measurements, electrical measurements, chemical
measurements, sonic measurements, and any combination thereof.
According to this embodiment the apparatus may comprise, for
instance: an extraction unit; an extraction unit and a nucleic acid
amplification unit; an extraction unit, a nucleic acid
amplification unit, and a detection unit. In each of these options
a reagent storage unit may be present in addition to the elements
of each option listed in the previous sentence. The extraction unit
allows a nucleic acid to be obtained from a sample processed by the
apparatus. The nucleic acid amplification unit allows a nucleic
acid obtained from the sample to be amplified (using, for instance,
PCR). The reagent storage unit comprises a reagent needed for, for
instance, extraction and/or amplification.
[0153] In order to have a wide spectrum of measurement
possibilities different types of sensors and detectors may be
installed within the device. Additionally it may be advantageous to
combine the already existing ultrasonic means for actuating or
treating the sample with the possibility to do sonic measurements.
The detection unit may be also part of the cartridge. In other
words optical readout, but also other detection labels e.g.
magnetic, electrical, electro-magnetic especially radio-frequency
applied techniques but also labelless methods are possible.
[0154] According to another exemplary embodiment of the invention,
the device further comprises a processor for coordinating a
treatment protocol, a data processor, a display and a user
interface.
[0155] It shall be noted that preferred embodiments of other
aspects of the present invention shall be considered as preferred
and disclosed embodiments with respect to the present aspect, too,
and vice versa.
[0156] According to another exemplary embodiment of the invention
the full solid coupler is made out of a polymer based material; and
wherein the polymer based material has a glass transition
temperature T.sub.g selected from the group comprising:
T.sub.g.gtoreq.-30.degree. C.; T.sub.g.gtoreq.-10.degree. C.;
T.sub.g.gtoreq.-5.degree. C.; T.sub.g.gtoreq.20.degree. C.;
T.sub.g.gtoreq.40.degree. C.; T.sub.g.gtoreq.60.degree. C.;
T.sub.g.gtoreq.80.degree. C.; T.sub.g.gtoreq.100.degree. C.;
T.sub.g.gtoreq.120.degree. C.; T.sub.g.gtoreq.130.degree. C.;
T.sub.g.gtoreq.140.degree. C.; T.sub.g.gtoreq.150.degree. C.; and
T.sub.g.gtoreq.160.degree. C.
[0157] It shall be noted, that the relevance of the glass
transition temperature of the material of the full solid coupler
gets more important the higher the intensity of the HiFu is. For
low intensity, for example when the input power P of the transducer
is smaller than 3 Watt the value of T.sub.g may not be that
relevant. This may be seen in FIG. 22. Medium intensity, P being
e.g. between 3 and 6 Watt, self enforced attenuation as described
above and hereinafter may play a more serious role which may
require a polymer with a sufficiently high T.sub.g. At high
intensities above e.g. 6 Watt the relevance of the choice of the
polymer based on his T.sub.g value even gets more important.
[0158] It may be necessary that at high intensity HiFu applications
at room temperature the T.sub.g may have to be above room
temperature (approximately 50.degree. C.).
[0159] It has been found that materials with relative high glass
transition temperature T.sub.g keep contrary to lower T.sub.g
materials during operation of the device and thus during the
acoustic energy transmission their low attenuation characteristics.
Thus an application of these low-attenuation high-T.sub.g materials
as full solid couplers enables very efficient transmission of
ultrasound intensities relevant for e.g. treatment of samples, e.g.
lysis of cells. Especially for HiFu applications as defined above
this is an advantageous effect realized by the invention. In other
words by using these materials a reduced power provided to the
source may be necessary to realize a certain HiFu power in the
focal region. Thus treatment and/or pretreatment functionalities
may be realized with a reduced power value. This may save energy
and costs. In other words the effect of self-enforced attenuation
of the coupling material may be avoided by the invention. The
attenuation per meter of the propagation path may thus be
reduced.
[0160] In order to provide for a better understanding of this
exemplary embodiment of the invention the following description of
the physical processes shall be noted:
[0161] Intrinsic to attenuation is that the coupler material
temperature may start increasing. Further the attenuation of
acoustic energy may also increase in parallel. This exemplary
embodiment of the invention now provides for materials that have
the advantage to keep a relatively low attenuation even when their
temperature starts increasing during e.g. HiFu operation in the MHz
range.
[0162] Examples for such materials may be polypropylene with
T.sub.g approximately -18.degree. C., epoxy with T.sub.g
approximately 60.degree. C. and silicons with T.sub.g approximately
60.degree. C., approximately 100.degree. C. and approximately
125.degree. C.
[0163] It has to be noted that a sufficiently high glass
temperature is related to the attenuation at the start of the test,
the ultrasound intensity, the thermal conductivity of the setup
(transport of heat generated) and the exposure time.
[0164] In other words the choice of the polymer with a certain
T.sub.g value depends on several parameters like the attenuation
value of the polymer at the beginning of the HiFu transmission
through the polymer as full solid coupler and thus before any
absorption or heat generation has started. Furthermore the applied
intensity or the power of the source determines that choice of the
polymer. Additionally the thermal conductivity of the surrounding
of the coupler is a parameter which influences the choice of a
polymer with a sufficiently high T.sub.g value. A high thermal
conductivity of the system around the coupler results in slower
temperature rise and lower maximum temperature, if HiFu is
sufficiently long exposed to reach equilibrium.
[0165] Especially for relatively high-intensity ultrasound
applications this may be particularly relevant. For example when
doing lysis with the HiFu energy within the sample the necessary
power may be relatively high. Thus for lysing methods using HiFu
this exemplary embodiment may be quite advantageous.
[0166] In other words the full solid coupler present in the
propagation between the source and the destination being the
cartridge has a reduced attenuation of the acoustic energy. In
addition to that tuned or matched impedances of the materials
transmitting the acoustic energy may be used, to minimize
reflection losses when passing material interfaces.
[0167] Thus the device provides for a complete dry coupling with
the possible following advantages: ease-of-use for the operator and
reduction of test turn-around-time, as a consequence other
applications run by less-skilled personal could be envisioned.
[0168] Polymers are a particular advantage class of materials to be
used as couplers, because of the rich variety of materials
available, the shape and dimensional design freedom, easy
replication and associated relative low costs. This may further be
described in detail in FIGS. 21 to 23.
[0169] According to another exemplary embodiment of the invention
wherein the polymer based material has been cured at a curing
temperature T.sub.c selected from the group comprising:
T.sub.c.gtoreq.20.degree. C.; T.sub.c.gtoreq.40.degree. C.;
T.sub.c.gtoreq.60.degree. C.; T.sub.c.gtoreq.70.degree. C.;
T.sub.c.gtoreq.80.degree. C.; T.sub.c.gtoreq.90.degree. C.;
T.sub.c.gtoreq.100.degree. C.; T.sub.c.gtoreq.110.degree. C.;
T.sub.c.gtoreq.120.degree. C.; T.sub.c.gtoreq.130.degree. C.;
T.sub.c.gtoreq.140.degree. C.; T.sub.c.gtoreq.150.degree. C.;
T.sub.c.gtoreq.160.degree. C.; T.sub.c.gtoreq.170.degree. C.; and
T.sub.c.gtoreq.180.degree. C.
[0170] The attenuation of the full solid coupler may further be
reduced ceteris paribus when the curing temperature of the polymer
based material during polymer fabrication is increased. This may
further be described in detail in FIGS. 21 to 23.
[0171] It has been found, that during the polymer fabrication,
which includes a curing process step, the curing temperature during
that curing process step at least partially determines the
transition glass temperature of the built polymer material. As
described above a sufficiently high T.sub.g value has certain
advantages for applications in a HiFu molecular device. Thus by
defining the curing temperature to a certain value a desired
T.sub.g value may be realized in the polymer. Such a process step
may be part of a method according to another exemplary embodiment
of the invention.
[0172] According to another exemplary embodiment of the invention,
a method for irradiating a sample with focused acoustic energy to
treat the sample is provided. Thereby the method comprises the
following steps: providing for an instrument, providing for a
cartridge, providing for a full solid coupler, providing for a
source for generating the acoustic energy, and inserting the
cartridge into the instrument. Furthermore the cartridge has a
chamber for receiving the samples and due to the inserting of the
cartridge into the instrument a complete dry coupling of the
acoustic energy between the source and the cartridge is provided.
The cartridge and the instrument are separable.
[0173] According to another embodiment of the invention an
instrument for irradiating a sample with focused acoustic energy to
treat the sample is presented. The instrument comprises a source
for generating the acoustic energy, a full solid coupler, wherein
the instrument is adapted to receive a cartridge containing the
sample, wherein the full solid coupler provides a complete dry
coupling of the acoustic energy between the source and the
cartridge, when the cartridge is inserted in the instrument;
wherein the cartridge and the instrument are separable and wherein
the instrument and the cartridge form a device according to one of
the above described embodiments.
[0174] This embodiment of the invention may be used with HiFu
acoustic energy in order to treat the sample with methods or
functionalities like mixing and/or lysing in e.g. one single
chamber. Furthermore the instrument may comprise a detector and an
excitation source that may both be for doing optical, electrical,
magnetic and/or mechanical measurements. Additionally a lens may be
comprised in the instrument.
[0175] In other words the dry coupling may be realized by the full
solid coupler that is part of the instrument. Before the presence
of the cartridge a complete dry propagation path from the source
through the full soil coupler is realized. By inserting the
cartridge into the instrument the whole dry propagation path
between the source and the sample is completed and the acoustic
energy may be transferred to the sample in order to treat the
sample.
[0176] According to another embodiment of the invention cartridge
for an instrument for irradiating a sample with focused acoustic
energy to treat the sample is presented, the cartridge comprising a
chamber for receiving the sample, a full solid coupler, wherein the
cartridge is adapted for being inserted in the instrument.
Furthermore the full solid coupler provides a complete dry coupling
of the acoustic energy between the source and the cartridge when
the cartridge is inserted in the instrument, wherein the cartridge
and the instrument are separable and wherein the instrument and the
cartridge form a device according to one of the above described
embodiments.
[0177] The full solid coupler may be permanently fixed to the
cartridge. But other solutions are possible. The source may for
example be part of the instrument. By inserting the cartridge into
the instrument the full solid propagation path between the source
and the sample is established.
[0178] Furthermore the source may be comprised by the cartridge.
Thus by inserting the cartridge into the instrument electrical
leads from the instrument are contacted with the source, in order
provide the source with electrical energy.
[0179] For the two before mentioned embodiments it shall explicitly
be noted, that the full solid coupler is arranged at the instrument
or at the cartridge in such a way, that the full solid coupler does
not have to form the whole propagation path by itself and other
additional dry coupling elements may be present. Nevertheless if it
is desired an exemplary embodiment of the invention may realize
this.
[0180] It shall further be noted that a computing unit may be part
of the instrument. It may be a separate unit in communication with
the instrument, or computing tasks may be distributed over computer
unit and instrument.
[0181] It shall further be noted that all computer program elements
mentioned above as exemplary embodiments of the invention might be
stored on a computing unit, which might also be part of an
embodiment of the present invention. This computing unit may be
adapted to perform or induce the performing of the steps of the
method described above. Moreover, it may be adapted to operate the
components of the above-described device. The computing unit can be
adapted to operate automatically and/or to execute the orders of a
user. Furthermore the computing unit can request the selection from
a user to process the input from the user.
[0182] The embodiments concerning computer program elements cover
both a computer program, that right from the beginning uses the
computer program element and a computer program that by an update
turns an existing program into a program that uses the
invention.
[0183] According to a further embodiment of the present invention,
a computer-readable medium is presented wherein the
computer-readable medium has a computer program element stored on
it which computer program element is described by the preceding or
following sections.
[0184] It may be seen as a gist of the invention that a consumable
cartridge being separable from the instrument generates a complete
dry coupling propagation path for focused acoustic energy when the
cartridge is inserted into the instrument. Thereby the dry coupling
reaches from the source generating the acoustic energy to the
sample.
[0185] It has to be noted that some of the embodiments of the
invention are described with reference to different
subject-matters. In particular, some embodiments are described with
reference to method type claims whereas other embodiments are
described with reference to apparatus type claims. However, a
person skilled in the art will gather from the above and the
following description that unless other notified in addition to any
combination or features belonging to one type of subject-matter
also any combination between features relating to different
subject-matters is considered to be disclosed within this
application.
[0186] The aspects defined above and further aspects, features and
advantages of the present invention can also be derived from the
examples of embodiments to be described hereinafter and are
explained with reference to examples of embodiments. The invention
will be described in more detail hereinafter with reference to
examples of embodiments but to which the invention is not
limited.
BRIEF DESCRIPTION OF THE DRAWINGS
[0187] FIG. 1 shows a schematic image of a device for irradiating a
sample with focused acoustic energy to treat the sample according
to an exemplary embodiment of the present invention.
[0188] FIG. 2 shows a schematic image of a cartridge having an
acoustic window according to an exemplary embodiment of the present
invention.
[0189] FIG. 3 shows a schematic image of a cartridge according to
an exemplary embodiment of the present invention.
[0190] FIGS. 4 to 8 show schematic images of sources of a device
according to an exemplary embodiment of the invention.
[0191] FIG. 9 shows a schematic image of several components of an
instrument according to an exemplary embodiment of the present
invention.
[0192] FIGS. 10 to 14 show exemplary overviews of possible
configurations of a device according to exemplary embodiments of
the present invention.
[0193] FIG. 15 shows a schematic image of electronic components
being used for a device according to an exemplary embodiment of the
present invention.
[0194] FIG. 16 shows a treatment protocol that is processed by a
device according to an exemplary embodiment of the present
invention.
[0195] FIGS. 17 to 19 show schematic images of devices generating
multi-focality to the sample according to an exemplary embodiment
of the present invention.
[0196] FIG. 20 shows a flow diagram representing a method according
to an exemplary embodiment of the present invention.
[0197] FIG. 21 shows a schematic image of a device for irradiating
a sample with focused acoustic energy to treat the sample according
to an exemplary embodiment of the present invention.
[0198] FIGS. 22 and 23 show a diagrams of results obtained with a
device for irradiating a sample with focused acoustic energy to
treat the sample according to an exemplary embodiment of the
present invention.
DETAILED DESCRIPTION OF THE INVENTION
[0199] Similar or relating components in the several figures are
provided with the same reference numerals. The view in the figure
is schematic and not fully scaled.
[0200] FIG. 1 shows a device 100 for irradiating a sample 101 with
focused acoustic energy to treat the sample according to an
exemplary embodiment of the present invention. It can clearly be
seen that the device has several components being an instrument
102, a cartridge 103, and a source 105 (shown only with dashed
lines) for generating the acoustic energy. Furthermore, a schematic
drawing of the propagation path 106 (dashed dotted lines) of the
acoustic energy starting at the source 105 and ending at the sample
101. Thereby the cartridge has a chamber 110 for receiving the
sample 101. Inside of the shown instrument 102 a full solid coupler
(not shown) 104 is provided in order to generate a propagation path
without non-fluidic matter. Thereby the source 105 and the full
solid coupler 104 are located inside of the instrument 102 and thus
cannot directly be seen. Furthermore the instrument 102 and the
cartridge 103 are adapted for inserting the cartridge into the
instrument wherein the cartridge and the instrument are separable.
It shall be noted that the hidden components like the source, the
lens, the full solid coupler and the acoustic window can be seen on
the following FIG. 9 showing an exploded view and FIG. 2
respectively.
[0201] Additionally a detection unit 111, e.g. a sensor, is shown
inside of the cartridge in order to do measurements on the sample
after or before a possible treatment by the focused acoustic
energy. Furthermore a processor for coordinating a treatment
protocol 112 is shown which is linked with the detection unit 111
and which is also connected to a display 114 and a data processor
113. The processor 112 for coordinating a treatment protocol is
connected to the device 100 and is further connected to the
detection unit 111. Thus the processor 112 is enabled to control
this complete-in result-out system in which in a fully automated
way a treatment of a sample by means of focused acoustic energy
especially by HiFu can be combined with analysis and measurements
as for example optical measurements, magnetic measurements, thermal
measurements, electrical measurements, chemical measurements, sonic
measurements and any combinations thereof.
[0202] Due to the use of HiFu and the corresponding short
wavelength (compared for example to known ultrasound applications
operating in the 20 kHz-100 kHz range) the size of focal region can
be decreased and thus a miniaturization of the whole molecular
device is possible. This is a highly important advantage of the
shown embodiment of the present invention with for example hospital
or lab requirements to have real small size systems because of the
very limited space available in these surroundings. Furthermore,
the combination of the functionalities treatment, pretreatment,
lysis and previously or subsequently done measurements may reduce
the costs and time of such a sample treating or molecular
diagnosis.
[0203] Additionally it may be possible to provide as such a device
100 with a multi-focality setup. Thereby the device generates at
least two different focal regions at the sample 101. This may be
done by at least two different sources, a single source and a
hybrid lens, or a single source with different roughness stones.
Furthermore a combination of these possibilities is also
possible.
[0204] Furthermore this device 101 may be used to reduce the
viscosity of a sample by means of the focused acoustic energy
especially by using HiFu.
[0205] In addition to that the device makes it possible to combine
in one single chamber 110 pretreatment and/or incubation and/or
lysis by means of focused acoustic energy originating from only one
single source 105. Especially a HiFu application is possible.
Thereby pretreatment and lysis may comprise different
functionalities that have been described in previous sections. This
may reduce costs and time of such a sample treating or molecular
diagnosis and also the space claimed of the device may be reduced
due to the integration of both functionalities into one chamber.
Furthermore the technical complexity of the device may be
reduced.
[0206] A pretreatment method or a lysis method may be processed or
carried out by means of the focused acoustic energy, especially by
HiFu and thus by the acoustic source or transducer generating the
HiFu spot at the position of the sample yielding to a pretreatment
and/or lysis of the sample. But also other devices that may be
integrated into the molecular diagnostic device and that are
necessary to carry out the method may generate the desired method.
For example an additional heating device, cooling devices, or
reagent applicator (dispenser) with supply lines may be integrated
in the molecular diagnostic device to cause incubation with an
additional reagent at elevated temperature.
[0207] A reagent may for example be lysozyme enzyme which may first
be mixed and subsequently incubated at 37.degree. C. Especially
mixing, circulation, liquefaction and homogenation may be done by
means of the irradiation of the sample with HiFu.
[0208] Furthermore also lysis of micro-organisms like e.g.
gram-negative and gram-positive bacteria, fungi and yeast may be
done by means of HiFu with the device 100 shown in FIG. 1. Lysing
may further comprise incubation of the sample with a reagent at
room temperature or elevated temperature. Reagents may for example
be GuHCl/prot K which is first mixed and subsequently incubated at
approximately 56.degree. C. and optionally cooled down to
environmental temperature or GuSCN which is first mixed and
subsequently incubated at approximately 70.degree. C. and
optionally cooled down to approximately 25.degree. C.
[0209] Optionally the chamber has at its outlet a filter or in its
outlet channel a filter to assure that debris is not transported to
the extraction functionality of the cartridge.
[0210] FIG. 2 shows an acoustic window 107 of the cartridge 103
wherein the acoustic window is made of a flexible material which is
shown as a plastic foil 108. It can be seen that the
circular-shaped acoustic window 107 that is shown in a bottom view
is covered by the plastic foil 108 being the interface medium that
may adapt itself to the shape of firstly the cartridge 103 and
secondly to a full solid coupler or source may be brought in
contact with the plastic foil directly on the shown surface 108.
115 shows the bottom part of the cartridge on which a flexible foil
is e.g. laser welded.
[0211] FIG. 3 shows the cartridge 103 with the chamber 110 in its
normal or working orientation which is a 180.degree. rotation
compared to FIG. 2. In other words, FIG. 2 shows the bottom part
115 of the cartridge with its bottom side and FIG. 3 shows the
cartridge with the bottom part 115 from the upper side. The shown
cartridge and foil clamp can then together as one unit be inserted
into the device 100 of FIG. 1 and can be pushed on top of the
instrument 102. This inserting process will form a propagation path
for transmitting the acoustic energy from the source 105 (shown
with dotted lines) in FIG. 1 to the sample 101 in FIG. 1.
[0212] FIG. 4 shows an example of a possible source used in a
device according to an exemplary embodiment wherein a source 105
and a coupler 104 is shown wherein the here shown example is a
polymer coupler.
[0213] FIG. 5 shows another example of a source creating the
focused acoustic energy especially HiFu wherein the source 105 may
be a piezo transducer and a metal lens 109 is fixed on top of that
for example flat transducer. Additionally a coupler 104 is provided
for example a polymer coupler.
[0214] In contrary to that FIG. 6 shows a polymer coupler
configuration in which a curved source 105 is combined with a
polymer coupler 104. In addition to that for example a lens may be
located on top of the polymer coupler being provided with another
for example polymer coupler on top of the lens to provide for an
efficient dry coupling towards the cartridge.
[0215] FIG. 7 shows a piezo configuration in which a flat piezo
transducer working as a natural focusing source 105 can be seen.
Additionally a very thin polymer layer is applied to modify the
roughness of the surface to promote efficient dry coupling. In
addition to that the electric leads are also shown.
[0216] FIG. 8 shows another possible configuration of the source
components in which a metal lens 109 is directly contacted to the
flat transducer working as a source 105. As will be later on seen
in FIGS. 10 to 14 any combination of these configurations is
possible which leads to a wide spectrum of applications.
[0217] FIG. 9 shows an exploded view of an instrument 102
comprising a heat sink 900, different housing rings 901 partially
building up the housing for the full solid coupler 104 that might
e.g. be a polymer based material or a solid gel, an additional ring
902. Furthermore the source 105 is shown as a piezo transducer.
Additionally the full solid coupler 104 is denoted with dotted
lines. These elements may be part of the instrument 102 and they
may build a receiving component that by inserting a cartridge on
top of the foil clamp 903 creates a propagation path that only
consists of non-fluidic matter. The elements 901, 902 and 903 are
part of the housing of the coupler, too. The housing is made such
that the height of the coupler could be modified by choosing number
of housing rings 901. The foil clamp 903 is clamped to the foil
(not depicted) which is used to cover the coupler.
[0218] FIG. 10 shows an overview of combinations of possibilities
to create dry coupling. Thereby the first row gives information
about the setup of the cartridge 103, the second row gives
information about the setup of the coupler 104, the third row gives
information about the setup of the lens 109 and the fourth row
gives information about the setup of the source or transducer 105.
It can be seen that five different configurations are shown as
examples. 1001 shows a solid gel coupler configuration wherein 1002
shows a metal lens polymer coupler configuration and 1003 describes
a polymer coupler configuration. 1004 describes a solution for the
dry coupling where a piezo only configuration (wherein the piezo
has a thin polymer layer to modify the roughness surface of the
transducer) is used and 1005 describes how a metal lens
configuration may be set up in order to reach dry coupling. 1001
shows that a source may be shaped like a lens and thus defines the
generation and the focusing of the acoustic energy. Furthermore
shown in column 1002 the lens may be physically combined for
example may be glued together with the solid coupler 104.
Furthermore the full solid coupler 104 may be directly attached to
the source 105 as shown in column 1003. But also a direct contact
between the cartridge and the piezo source is possible as shown in
1004. Additionally the metal lens configuration describes that at a
curved shaped source 105 can be attached a biconcave shaped lens
e.g. a metal lens.
[0219] Other setup possibilities may be shown in the detailed
overviews 1100 within the FIGS. 11, 12, 13 and 14. These overviews
are more detailed than FIG. 10 because two additional rows are
inserted in order to distinguish between the fact whether a
component is part of the cartridge, is part of the source (which
means is part of the instrument) or is a physically separated
component.
[0220] It shall explicitly be noted that any shown and described
component may be part of the transducer, of the cartridge or may be
a physically separated component. In addition to that any
combination of components may be used in order to separate
different functionalities. For example a thin foil, having a high
flexibility may be used to adapt the shape of a transducer. In
combination with a full solid coupler having less flexibility but
lower attenuation than the foil, this corresponds to the separation
of the functionalities attenuation and flexibility. This may lead
to an advantageous combination of different components to achieve
efficient dry coupling.
[0221] Row 1101 describes, if there is an entry, that the full
solid coupler is part of the cartridge. In contrary to that 1102
describes the fact that the full solid coupler is part of the
source and thus part of the instrument. Also both possibilities may
be arranged at an device simultaneously. As a third possibility
1104 describes that the full solid coupler is a physically
separated component being inserted into the propagation path. Again
it can be seen that a combination of lens and source 1103 may be
provided. As can be seen from FIGS. 11 to 14 a huge variety of
setup possibilities for the dry coupling of the device using for
example HiFu is possible.
[0222] FIG. 15 shows exemplary electronic components 1500 being
used to generate the focused acoustic energy. Thereby a possible
function generator, a power amplifier, a scope and an ultrasonic
transducer are connected together in order to create the acoustic
field. After having focused the emitted acoustic energy it impinges
the sample and causes different sono-chemical or sono-physical
reactions. This is the treatment of the sample caused by device. In
other words FIG. 15 shows a configuration of a lab setup to
generate and investigate the setup performance. An industrial
device may not include a scope and the function generator and the
amplifier may be embodied in specific and custom made
electronics.
[0223] FIG. 16 shows a possible treatment protocol for applying
pretreatment and lysis in one single chamber by only one single
source. Treatment protocol 1600 has several steps for example the
protocol starts with a HiFu pretreatment of the sample 1603,
subsequently a mixing 1604 is applied to the sample wherein
afterwards an incubation with different matter 1605 is possible.
Subsequent additional mixing and incubation steps are possible.
These different functionalities created or caused by the acoustic
energy due to sono-chemical or sono-physical interactions are all
part of the pretreatment 1601. Subsequently a lysis 1602 is
possible within the same single chamber and can be caused by the
same single source that has been processed the pretreatment. As
possible steps mixing and incubations may be mentioned. But also
special HiFu lysis 1606 and additional filter steps 1607 are
possible. Thereby reference sign 1608 describes any sample with a
target material to be detected, e.g. feces, blood, urine, sputum,
BAL, CSF, tissue, swab or brush. Furthermore a first pretreatment
reagent (e.g. chemical compound(s) and/or enzyme(s)) is shown with
1609. A second pretreatment reagent (chemical compound(s) and/or
enzyme(s) is shown with reference sign 1610 and 1611 depicts a
third pretreatment reagent (chemical compound(s) and/or enzyme(s)).
A first lysis reagent (chemical compound(s) and/or enzyme(s)) is
shown by 1612. 1613 shows an extraction reagent, e.g. to prepare
for DNA binding on silica. The shown figure is only an exemplary
embodiment and a filter does not have to be inside the lysis
chamber.
[0224] FIG. 17 shows a multi-focality setup 1700 of the device
according to another exemplary embodiment of the invention. It can
be seen that the cartridge 103 having a chamber 110 with a sample
101 also features the possibility to have an air volume 1701 above
the sample. Furthermore two different sources 105 are applied in
the setup in order to generate a first focal region 1702 and 1703
showing a second focal region. Furthermore the acoustic window of
the cartridge should have a low attenuation and minimal thickness
to avoid heating of the material and to realize a high intensity in
the focal regions. For mass production an injection moldable
polymer is preferred. It may be preferred that no contact is made
of the focal regions with the walls of the chamber. At high
intensities this may result in melting of the wall. It may further
be desired that the transducer with the large focal zone 1702 is
placed opposite to the air volume 1701. This results in optimal
mixing and circulation and may have lower risk on melting the
chamber wall.
[0225] FIGS. 18a and 18b show multi-focality of the device working
for example in the HiFu range, may be generated by only one single
source. Thereby FIG. 18a shows a multi-focality setup 1700 with a
hybrid lens 1800 having a first emitting zone 1801 and a second
emitting zone 1802 and a third emitting zone 1803. It is also
possible that in a concentric setup the first and the third
emitting zones are equal. It can further be seen, that in the
sample 101 three different focal regions 1804 to 1806 are
generated. In a concentric setup it is thus the case that 1804 and
1806 describe the same focal region having a ring-like shape around
the second focal region 1805.
[0226] It can be seen that the source 105 may be of a flat shape
and the hybrid lens 1800 is attached to the source.
[0227] FIG. 18b shows a multi-focality setup 1700 wherein the
hybrid lens 1800 has got a shape that is adapted to the shape of
the curved source 1500. In FIG. 18b the hybrid lens has three
emitting zones and three focal regions originating from the three
emitting zones. The different emitting zones may consist of
different focusing material. For example, the outer material
forming zone 1801 and 1803 may be of moderately focusing outer
material wherein the inner material forming the zone 1802 may be a
highly focusing material. The segmented lens 1800 thus comprises
highly focusing material and moderately focusing material. This may
be the case for FIG. 18b. These different focal regions may enable
a user to process different functionalities like mixing and lysing
simultaneously by only using one single source. This may reduce the
times of for example a molecular test and furthermore costs and
space requirements may be reduced as only one single source is
needed. Additionally technical problems and maintenance costs are
reducible.
[0228] Thereby the distribution of the differently focusing
materials can be adapted the desired treatment, lysis or analyzing
application. Thus no specific material distribution within the
hybrid lens or segmented lens is excluded by this exemplary
embodiment of the invention.
[0229] The following paragraph relates to modeling of a combination
comprising a flat transducer and a curved lens to verify the hybrid
lens concept. A possible setup may be for example a high impedance
material like for example aluminum, a low impedance material like
polypropylene taken as a lens material, a lens radius and internal
diameter of the chamber like for example 8 mm and polypropylene is
taken as chamber wall material with a thickness of 0.5 mm, a fluid
height is 35 mm and the frequency for the modeling is 1 MHz and
prescribed pressure piezo is 1.000 Pa. The results of the modeling
disclosed that the maximum pressure along the central axis of
symmetry remains at a very constant high level when going from a
complete high impedance material (aluminum) to increasing segment
sizes of low impedance material like polypropylene. In other words,
the pressure remains at the level sufficient high to obtain lysis.
Secondly the results revealed that a minimum and maximum pressure
conditions are created outside the central axis of the chamber when
the polypropylene segment size is sufficiently large to create
mixing. Effective working of the hybrid construction is achieved
when the high index material (aluminum for instance) is typically
between 1/5 and 1/2 of the total lens when a low index material is
a low dissipation plastic. Thus a hybrid lens is an option to
generate multifocal acoustic energy especially multifocal HiFu from
a single piezo element. This solution could be used for HiFu across
dry interfaces as well as for liquid or hydrogel coupling and
direct contact with the fluid.
[0230] FIG. 19 shows a multifocal setup 1700 wherein a source 105
has different surface roughness zones. 1903 shows a top view of the
circular source 105 having a first surface roughness zone 1904 and
a second surface roughness zone 1905 yielding to multi-focality. It
can be seen that the first focal region 1900 and the second focal
region 1901 are different from each other. Here the third focal
region 1902 is the same as the first focal region 1900 because the
second roughness zone 1905 is a ring-shaped surface that yields to
a ring-shaped focal region 1900 and 1902 around the second focal
region 1901. Due to different roughnesses of the surfaces a
different coupling to material transmitting the acoustic energy is
given. Therefore, different roughnesses yield in different focal
regions.
[0231] It shall explicitly be noted that the multi-focality due to
different surface roughnesses may not be used with the dry coupling
features of the present invention and may be applied independently
on a device for irradiating a sample with multi focused acoustic
energy to treat the sample.
[0232] For example, in the range of 1 to 2 MHz the effect may be
moderate for a roughness of 10 .mu.m and may be significantly
higher for a roughness of 50-80 .mu.m. Thus, a curved transducer
with rough and smooth segments is an option to generate multifocal
HiFu from a single piezo element. Compared to different solutions
with lenses or a plurality of sources this embodiment may be
simpler.
[0233] FIG. 20 shows a flow diagram describing a method for
irradiating a sample with focused acoustic energy to treat the
sample wherein the following steps are comprised and for an
instrument S1, providing for a cartridge S2, providing for a full
solid coupler S3, providing for a source for generating the
acoustic energy S4. Furthermore inserting the cartridge into the
instrument S5 wherein the cartridge has a chamber for receiving the
sample and wherein due to the inserting of the cartridge into the
instrument a complete dry coupling of the acoustic energy between
the source and the cartridge is provided. Furthermore the cartridge
and the instrument are separable.
[0234] FIG. 21 shows a schematic drawing of an instrument device
comprising a transducer 105, a full solid coupler 104, a cartridge
103 having a chamber 110 for a sample to be treated with e.g. HiFu
by the instrument 102. The bottom 2100 of the cartridge has an
acoustic window made out of a foil.
[0235] FIG. 22 shows a diagram 2200 in which the advantages of a
full solid coupler with a sufficiently high glass transition
temperature T.sub.g are illustrated. It can be seen from the graphs
2203-2207, that a full solid coupler with higher glass transition
temperature T.sub.g provides for less attenuation of the ultra
sound energy within the full solid coupler. These results shall be
describe in detail hereinafter.
[0236] The x-coordinate 2201 depicts the input power that is
provided to the source 105 (not shown) which generates the acoustic
energy e.g. the HiFu. The y coordinate depicts the so called
clipping time. This is the time between the source generating e.g.
the HiFu is switched on and the complete disappearance of the
fountain (clipping). This fountain generation has been described
above. It is created by the HiFu waves and is used to reduce the
power threshold at which cavitation in the sample sets in. The
fountain is consisting of the sample material (e.g. a liquid). As
the generation of such a fountain depends on the acoustic energy
that is transmitted through the full solid coupler to the sample
the disappearance of the sample means a reduction of transmitted
acoustic energy. Different materials with different glass
transition temperatures are observed within the test of the results
shown in FIG. 22.
[0237] In other words clipping is taken as a measure for the
development of the attenuation or absorption with time of the
observed full solid coupler material. Results for a variety of
materials and thicknesses are presented in FIGS. 22 and 23.
[0238] Thereby FIG. 22 shows results from a 3 mm thick silicon 601
coupler 2203 having a glass transition temperature of 60.degree. C.
2204 depicts the results from a 3 mm thick full solid coupler made
out of epotek 301 having a glass transition temperature T.sub.g of
approximately 60.degree. C. 2205 depicts the results of a 6 mm
thick silicon 601 coupler having a glass transition temperature of
60.degree. C. 2206 depicts the results of a full solid coupler that
is 1 mm thick and made out of polypropylene (PP) having a glass
transition temperature T.sub.g of approximately -18.degree. C. 2207
depicts the results of a full solid coupler made out of 5 mm thick
epotek 301 having a glass transition temperature of approximately
60.degree. C. All examples have cure temperatures of 60 C except
PP.
[0239] FIG. 22 shows that PP is even at moderate intensity a rather
poor performer. Attenuation of both epoxy and silicone increases as
expected with thickness of the full solid coupler. Attenuation of
epoxy is for silicon lower than for epoxy. For all of these high
T.sub.g materials clipping is observed for continuous input power
<6 Watt. This power may be insufficient for sample treatment.
Thus for broad treatment possibilities of a molecular diagnostic
device the invention provides for sufficiently high T.sub.g
polymers.
[0240] Additional experiments have disclosed that firstly the
observed phenomena is not due to a change over time of the
transducer. Secondly the effect may be reversible (if the material
is not exposed to burn-through intensities). After about 1 min the
material is returned to its original state and the experiment could
be repeated. This observation suggests a temperature-material
property relationship.
[0241] FIG. 23 shows a diagram 2300 in which the effect of the
curing temperature of the polymer based material used as a full
solid coupler is shown. X-coordinate 2301 depicts the input power
and y-coordinate 2302 depicts the time to failure i.e. the clipping
time. 2303 to 2306 depict the graphs of the different full solid
coupler. 2303 depicts the result of a full solid coupler with the
curing temperature T.sub.c is 100.degree. C., 2304 depicts T.sub.c
is 125.degree. C., 2305 depicts T.sub.c is 60.degree. C. and 2306
also depicts the results of a coupler with T.sub.c is 60.degree. C.
In other words FIG. 23 shows that the effect attenuation is also
dependent on the curing temperature. With increasing curing
temperature the clipping time increases significantly. An exemplary
embodiment of the invention uses this advantage. In other words in
general a higher curing temperatures T.sub.c directly translates
into a higher glass transition temperature T.sub.g.
[0242] Additional experiments with the material cured at 60.degree.
C. have shown that:
[0243] Firstly fountain disappeared if water of 80.degree. C. or
more is used. Secondly with a duty cycle of 20% the clipping time
shift to >120 seconds for peak power between 0 W and 65 W
(average power 13 W). For a peak power of 90 W (average power 16 W)
the clipping time has reduced to 10 seconds.
[0244] Possible exemplary equipment devices for these test may be
the following: PM5193 Programmable Synthesizer/function generator
0.1 mHz-50 MHz, Amplifier: ENI 240L Power Amplifier 50 dB 20 kHz-10
MHz or AR worldwide KAA204 RF Power Amplifier 50 dB 0.5-100 MHz 200
W, Tektronix TDS3014: Four Channel Color Digital Phosphor
Oscilloscope; Agilent 4395A: 10 Hz-500 MHz/10 Hz-500 MHz/10 kHz-500
MHz Network/Spectrum/Impedance Analyzer and HiFu piezo transducer:
JR20/60 supplied by Dongfang Jinrong.
[0245] In the claims the word "comprising" does not exclude other
elements or steps, and the indefinite article "a" or "an" does not
exclude a plurality. Reference signs shall not limit the scope of
the claims.
LIST OF REFERENCE NUMERALS
[0246] 100 Device [0247] 101 Sample [0248] 102 Instrument [0249]
103 Cartridge [0250] 104 Full solid coupler [0251] 105 Source
[0252] 106 Propagation path [0253] 107 Acoustic window [0254] 108
Flexible material [0255] 109 Lens [0256] 110 Chamber [0257] 111
Detection unit [0258] 112 Processor for coordinating a treatment
protocol [0259] 113 Data processor [0260] 114 Display [0261] 115
Bottom part [0262] 900 Heat sink [0263] 901 Housing rings [0264]
902 Additional ring [0265] 903 Foil clamp [0266] 1000 Overview of
combination possibilities to create dry coupling [0267] 1001 Solid
gel coupler configuration [0268] 1002 Metal lens polymer coupler
configuration [0269] 1003 Polymer coupler configuration [0270] 1004
Piezo only configuration (wherein the piezo has a thin polymer
layer to modify roughness surface) [0271] 1005 Metal lens
configuration [0272] 1100 Detailed overviews of combination
possibilities to create dry coupling [0273] 1101 Row describing
that the full solid coupler is part of the cartridge [0274] 1102
Row describing that the full solid coupler is part of the
instrument [0275] 1103 Component combining the functionality of a
lens and a source (curved source) [0276] 1104 Row describing that
the full solid coupler is a physically separated component [0277]
1500 Electronics being used to generate the focused acoustic energy
[0278] 1600 Possible treatment protocol for applying pretreatment,
incubation and lysis in one single chamber by one single source
[0279] 1601 Pretreatment part of the protocol [0280] 1602 Lysis
part of the protocol [0281] 1603 HiFu pretreatment [0282] 1604
Mixing [0283] 1605 Incubation [0284] 1606 HiFu lysis [0285] 1607
Filtering [0286] 1608 Sample with a target material to be detected
[0287] 1609 First pretreatment reagent [0288] 1610 Second
pretreatment reagent [0289] 1611 Third pretreatment reagent [0290]
1612 First lysis reagent [0291] 1613 Extraction reagent [0292] 1700
Multi-focality setup [0293] 1701 Air volume above the sample [0294]
1702 First focal region [0295] 1703 Second focal region [0296] 1800
Hybrid lens [0297] 1801 First emitting zone of the hybrid lens
[0298] 1802 Second emitting zone of the hybrid lens [0299] 1803
Third emitting zone of the hybrid lens [0300] 1900 First focal
region [0301] 1901 Second focal region [0302] 1902 Third focal
region [0303] 1903 Top view of source 105 with different
roughnesses zones [0304] 1904 First roughness zone of the source
[0305] 1905 Second roughness zone of the source [0306] S1 Providing
for an instrument [0307] S2 Providing for a cartridge [0308] S3
Providing for a full solid coupler [0309] S4 Providing for a source
for generating the acoustic energy [0310] S5 Inserting the
cartridge into instrument
* * * * *