U.S. patent application number 14/814249 was filed with the patent office on 2016-01-07 for fluidic interfacing system and assembly.
The applicant listed for this patent is Spinomix, S.A.. Invention is credited to Amar Rida, Thierry Varidel.
Application Number | 20160001284 14/814249 |
Document ID | / |
Family ID | 44628036 |
Filed Date | 2016-01-07 |
United States Patent
Application |
20160001284 |
Kind Code |
A1 |
Rida; Amar ; et al. |
January 7, 2016 |
Fluidic Interfacing System and Assembly
Abstract
A fluidic assay system assembly comprising: (a) A disposable
fluidic cartridge (1) comprising at least one reaction chamber (3)
connected to a network of fluidic channels (2) with at least one
inlet channel and one outlet channel. The said inlet and outlet
channels end at the down side of the fluidic cartridge with at
least two connecting pores (4), (4'); (b) A disposable vessel (5)
comprising a connection tube 22 immersed in a sample container (6)
and ended at the cap of the vessel with an external connection pore
(7) ; (c) A fluidic manifold (8) that is interdependent with the
bulk system (12) comprising a fluidic network connected (9) to
active fluidic parts (10), (11). The said channel network ends at
the top side of the fluidic manifold with at least one connecting
pore (13). Wherein the first and the second pores of the fluidic
cartridge are interfaced by direct physical contact with the sample
container and the manifold pores, respectively.
Inventors: |
Rida; Amar;
(Chavannes-Remens, CH) ; Varidel; Thierry;
(Ecublens, CH) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Spinomix, S.A. |
Lausanne |
|
CH |
|
|
Family ID: |
44628036 |
Appl. No.: |
14/814249 |
Filed: |
July 30, 2015 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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13700644 |
Jun 26, 2013 |
9132422 |
|
|
PCT/IB2011/052440 |
Jun 3, 2011 |
|
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14814249 |
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Current U.S.
Class: |
422/527 |
Current CPC
Class: |
B01L 2200/0689 20130101;
B01L 2200/027 20130101; B01L 3/50825 20130101; B01L 2300/0672
20130101; B01L 3/502 20130101; B01L 2400/0655 20130101; B01L
2300/0887 20130101; B01L 2400/0487 20130101; B01L 2300/06 20130101;
B01L 2300/123 20130101; B01L 3/502715 20130101; G01N 33/54326
20130101 |
International
Class: |
B01L 3/00 20060101
B01L003/00 |
Foreign Application Data
Date |
Code |
Application Number |
Jun 3, 2010 |
CH |
00885/10 |
Claims
1.-11. (canceled)
12. A fluidic cartridge used for extracting target biomolecules or
particles from a crude sample, the cartridge comprising: a. at
least one structure top layer containing: i. a reaction chamber
with a solid support that is designed to capture the said target
biomolecules, ii. a first inlet and outlet channels that are in
fluid communication with the said reaction chamber and that will be
used to bring the sample it and out the reaction chamber, and iii.
a second inlet and outlet channels connected to the said reaction
chamber and that will be used for eluting the purified biomolecules
wherein the second inlet and outlet channels are diverging branch
of the first inlet and outlet channels.
13. The fluidic cartridge according to claim 12, wherein the solid
support is constituted by magnetic particles.
14. (canceled)
15. The fluidic cartridge according to claim 12, wherein the said
first inlet channel is in fluid communication with the sample
vessel.
16. The fluidic cartridge according to claim 12, wherein the said
second inlet channel is in fluid communication with a recovery
sample vessel.
17. The fluidic cartridge according to claim 12, wherein the
reaction chamber is further in fluid communication with a fluidic
network channel that brings reagents in the said reaction
chamber.
18. The fluidic cartridge according to claim 12, wherein the
cartridge further comprises a closing down layer composed from a
flexible material and comprising connection pores associated to the
ends of said inlet and outlet channels.
19. The fluidic cartridge according to claim 15, wherein in
processing, the sample is aspirated from the sample vessel, in
fluid communication with the first inlet channel, into the reaction
chamber through the first outlet channel.
20. The fluidic cartridge according to claim 16, wherein in
processing, the target biomolecules or particles are eluted into
the recovery sample vessel, in fluid communication with the second
outlet channel, from the reaction chamber by pouching air through
second inlet channel.
21. The fluidic cartridge according to claim 12, which comprises at
least one optical sensor positioned in one side of said reaction
chamber.
22. The fluidic cartridge according to claim 13, wherein the
magnetic particles are manipulated and mixed using at least two
electromagnetic poles face each other across the reaction chamber,
and wherein the said magnetic poles are actuated by: a) applying
magnetic field sequences having polarity and intensity that vary in
time from the electromagnetic poles, wherein said magnetic field
sequences break or inhibit the particle aggregates and maintain the
particles in suspension as a fog of particles in relative dynamic
movement; and b) combining the magnetic fields from different
magnetic poles in a sequence to induce displacement of the fog of
particles across the reaction chamber, wherein the fog of particles
occupies substantially the whole reaction chamber volume.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a Continuation of U.S. patent
application Ser. No. 13/700,644, which is a national stage
application, filed under 35 U.S.C. .sctn.371, of International
Application No. PCT/1B2011/052440, filed Jun. 3, 2011, which claims
the benefit of and priority to Swiss Patent Application No.
00885/10, filed Jun. 3, 2010. The contents of each application are
incorporated by referenced in their entirety.
FIELD OF THE INVENTION
[0002] The invention relates to a system for conducting an
automated assay on a specimen which contains specific biological or
chemical substances that need to be detected. More specifically,
the invention discloses a device that includes a fluidic cartridge
and the interfacing of the said cartridge with the sample and the
different reagents used in performing such assay.
BACKGROUND OF THE INVENTION
[0003] The testing of diverse sample types derived from human,
animal, plant sources, food and environmental samples plays a
crucial role in modern medical diagnosis and treatment, forensic
medicine, food safety, industrial processing among many other
fields. However, such analyses are very often related to complex
processes that relate to labor-intensive, chemical, biological and
physical steps on a fluid sample that will end in the detection of
specifically targeted molecules or analytes using optical,
electrical and biochemical procedures. In the current
state-of-the-art, sample analysis steps remain mainly dominated by
complex, large and expensive "robotic" instruments operated by
expert technicians in centralized laboratories. Consequently, any
technology that would automate the complex reaction and sample
processing steps while making them more affordable and less
space-consuming would address unmet needs for simple,
cost-effective assaying solutions.
[0004] As an emerging alternative to the robotic platforms, fluidic
or microfluidic technologies open new perspectives in assay
processing and automation. In relation to recent developments in
molecular biology, nanotechnology and optics, (micro)-fluidic based
systems comprehend indeed the potential of providing integrated
solutions, where all steps from sample preparation and assay
processing to signal amplification and detection of multiple
targets will be integrated in a fully automated, compact
cartridge.
[0005] A typical example of a commercially available
cartridge-based solution is the
[0006] GeneXpert molecular diagnostics platform from Cepheid (CA,
USA) (cepheid.com) which realized an advance in fully automated
molecular testing from sample input to result reporting. As for
instance disclosed by the U.S. Pat. No. 6,893,879, U.S. Pat. No.
6,664,104, U.S. Pat. No. 6,818,185 and U.S. Pat. No. 6,783,736, the
Cepheid system demonstrates the integration of micro-fabricated
chips and other miniaturized fluidic or analytical components in a
cartridge type where steps from the separation of a desired analyte
from the original sample fluid sample to assay processing and
target detection are being performed.
[0007] Beyond the integration capabilities, one fundamental issue
of the development of a micro-fluidic based system is the
interfacing of the fluidic part which is substantially small and
compact as compared to the relatively large macro-environment, as
defined by the user samples, reagents and sensing elements.
[0008] With this respect, the international Pat. Application WO
2008/030433 for instance describes the use of a cartridge, which is
adapted to contain samples and reaction fluids to interface with a
micro-fluidic chip for use for DNA analysis tests and other assays
performed within the micro-fluidic chip. The microfluidic interface
is assured through access ports in connection with microfluidic
channels and located on the top side of the associated
micro-fluidic chip. The reagents and samples contained in external
chambers within a fluidic cartridge are dispersed into the
microfluidic chip through nozzles that will be brought in
communication with the access ports on top of said microfluidic
chip. Within the same spirit, WO 2010/118427 discloses a fluidic
interface device that includes a cartridge with microfluidic
configuration that is in fluid communication with a microfluidic
chip through contact pores.
[0009] The state-of-the-art coupling and interfacing of the
micro-fluidic based system with the external environment systems,
are facing however a real challenge: The integration of maximum
functionalities within the micro-fluidic cartridge which leads to a
complex and costly "disposable" cartridge or to lower integration
properties at the cartridge level resulting in a more complex
interfacing device that practically end in complex robotic
platforms. The optimal balance between the cartridge
complexity/simplicity versus the corresponding interfacing system
simplicity/complexity is still an open mostly unresolved issue.
[0010] In knowledge of these shortcomings the current invention
concerns a system for conducting automated assays within a fluidic
cartridge and its interfacing device on a sample containing
specific biological or chemical substances that need to be
detected. This disclosed system overcomes various limitation and
constraints by assuring the simplicity of both the disposable
cartridge and its interfacing system.
SUMMARY OF THE INVENTION
[0011] The present invention provides a system for the automated
procedure of bio-chemical assays that includes: [0012] a. A
disposable fluidic cartridge comprising at least one reaction
chamber connected to a network of fluidic channels with at least
one inlet and one outlet channels, wherein the said inlet and
outlet channels end at the down side of the fluidic cartridge with
at least two connecting pores; [0013] b. A disposable vessel
comprising a connection tube immersed in a sample container and
ended at the cap of the vessel with an external connection pore;
[0014] c. A fluidic manifold that is interdependent to the bulk
system comprising a fluidic network connected to active fluidic
parts, wherein the said channel network ends at the top side of the
fluidic manifold with at least one connecting pore; and wherein the
first and the second pores of the fluidic cartridge are interfaced
by direct contact with the sample and the manifold container pores
respectively.
[0015] The key advantage of this invention is the simplicity of the
automated system in managing the sample as well as the different
reagents that will be used in any assay process.
[0016] This simplicity is first translated by the fluidic cartridge
design that can be composed from plastic molded parts preferably
comprising of a structured layer with fluidic structures and of a
sealing layer. In a preferred realization of the invention, the
closing-down layer is composed of an elastomeric material which in
practice will serve as an interface to seal the connection of the
fluidic cartridge pores with the respective pores of the disposable
vessel and fluidic manifold.
[0017] Accordingly, the present invention discloses a fluidic
cartridge for assaying target biomolecules or particles from a
crude sample, the cartridge comprising of: [0018] d. at least one
structure top layer containing: [0019] i. a reaction chamber with a
solid support that is designed to capture the said target
biomolecules, [0020] ii. a first inlet and outlet channels that are
in fluid communication with the said reaction chamber and that will
be used to bring the sample it and out the reaction chamber, [0021]
iii. a second inlet and outlet channels connected to the said
reaction chamber and will be used for eluting the purified
biomolecules, [0022] iv. wherein the second inlet and outlet
channels are diverging branch of the first inlet and outlet
channels, [0023] e. a closing down layer composed from a flexible
material and comprising connection pores associated to the ends of
the said inlets and outlets channels,
[0024] Further, the simplicity of the automated system is realized
by handling the sample to be assayed within a disposable sample
vessel container from a traditional laboratory sample collection
tube with a pierceable cap. To assure the connectivity with the
fluidic cartridge a syringe adapter is inserted into the tube. Such
tubes will be preferably used for containing the starting sample
that will be assayed and thereby avoid the complexity induced by
the integration of sample storing directly into the fluidic
cartridge.
[0025] Further, the simplicity of the automated system is realized
by a fluidic manifold that comprises fluidic network channels and
active fluidic elements like valves and pumps mounted on the said
manifold. The fluidic manifold according to the invention is
further characterized by the fact that it is part of the bulk
system. Fluidic connectivity of the fluidic manifold with the
disposable fluidic cartridge is assured through connection pores
disposed on the top side of the manifold and that will be directly
in contact with the fluidic cartridge and the pores reflectively.
Furthermore, the fluidic manifold comprises connection pores that
will be in a fluidic communication with the diverse assaying
reagents. The latter will be transported through the fluidic
manifold network channels to be thereafter injected into the
fluidic cartridge through a specifically dedicated pore. To avoid
cross contamination, the manifold fluidic network channels are
divided into a first network segment of channels specifically
designed to handle the sample to be assayed and a second segment of
channels specifically designed to handle the reagents and wherein
the said first and second segment of channels are fluidically
disconnected from each other.
[0026] In summary, the current invention discloses a fluidic system
assembly for conducting bio-assays comprising of: [0027] (1) A
fluidic cartridge plastic element in which the assay will be
conducted. Without any active element integration, this disposable
element can be easily manufactured using standard plastic injection
molding and assembling techniques. Designed for specific assays,
the cartridge can further include assay specific reagents as
specific affinity or detection reagents preferably in a lyophilized
format. [0028] (2) A sample container and handling vessel that must
be also disposable to avoid any cross-contaminations. Rather than
to be directly integrated into the fluidic cartridge, the sample
container according to the invention is a standard laboratory tube
with a pierceable cap. To assure the fluidic connectivity,
preferably, a syringe adapter is inserted in the tube through the
said pierceable cap. [0029] (3) A fluidic manifold that contains
the system active elements (valves, pumps, actuation and sensing
elements) will be integrated. This part of the system is not
disposable and will be considered as part of the bulk of the
system. The fluidic manifold can also directly integrate the
generic reagents handling to avoid cross contamination issues.
[0030] (4) The manifold as well as the sample containers will be
directly interfaced with the fluidic cartridge through the
respective pores disposed at the down layer of the said fluidic
cartridge. The fluidic manifold can also serve as support to
receive the sample vessels. The microfluidic chip will be on the
top of the fluidic manifold and the sample vessel to provide the
full fluidic sealed assembly.
BRIEF DESCRIPTION OF THE DRAWINGS
[0031] The objects and features of the present invention are set
forth with particularity in the appended claims. The present
invention, both as to its organization and manner of operation,
together with further objects and advantages, may best be
understood by reference to the following description, taken in
connection with the accompanying drawings, wherein
[0032] FIGS. 1 is a schematic representation of the fluidic
assembly and samples interfacing according to a preferred
embodiment of the invention.
[0033] FIGS. 2 is a schematic representation of the fluidic
cartridge according to a preferred embodiment of the invention.
[0034] FIG. 3 shows a schematic view of a system realization that
includes the key features of the invention. The system design,
components and the connectivity of such components are shown in
accordance with the preferred embodiments of the invention.
[0035] FIG. 4 shows a schematic representation of the system of the
FIG. 3 after the different components of the system have been
assembled.
[0036] FIG. 5 shows a schematic representation of the system of the
FIG. 4 with an emphasis of the disposable vessels and the fluidic
manifold assembly according to a preferred embodiment of the
invention.
[0037] FIG. 6 is a schematic representation of the disposable
vessels according to a preferred embodiment of the invention.
[0038] FIG. 7 is a schematic representation of the liquid position
sensing which comprises a fluidic cartridge with a reaction chamber
and at least one optical sensor positioned on one side of the said
reaction chamber.
[0039] FIG. 8 is schematic representation of the liquid position
sensing method according to the invention.
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS
[0040] The main purpose of the present invention is an automated
system for sample handling, preparation and assaying using a
disposable of a simple designed and low cost fluidic cartridge.
Furthermore, the invention discloses an interfacing system of the
said cartridge with the sample to be assayed and also with the
reagents used in performing such assay. Another main attainable
objective of the present invention is a fully automated device for
biological liquid sample and reagent processing in a microfluidic
using the said fluid cartridge and the related interfacing
system.
[0041] In general, the microfluidic environment of the invention
concerns devices typically designed at a scale suitable to analyze
micro-volumes preferably in the range 0.1 ml to 500 .mu.l. However,
for one of the major applications of the invention large sample
sizes are used to concentrate specific biomolecules or biological
cells or particles in the device to a small volume for subsequent
analysis. The microscale flow channels and wells have preferred
depths and widths in the order of 0.05-1 mm. The "reaction chamber"
that is part of the microfluidic network as used herein refers to
chambers with a cavity that have a volume in the range of 1 .mu.l
to 1 ml and preferably in the range 10 .mu.l to 200 .mu.l. However,
for many applications, larger "mesoscale" dimensions in the scale
of millimetres may be used. Similarly, geometry features often will
have larger dimensions than the microchannels, in the scale of 1-10
mm width and 1-5 mm depth.
[0042] The disclosed system and devices herein can be applied to
perform complex assays used in various testing laboratories and
clinical procedures. Such procedures can include but are not
limited to extraction, purification and concentration of target
molecules or particles from a wide range of target substances in
biological samples. Examples of target substances are cells, cell
components, cell subpopulations (both eukaryotic and prokaryotic),
bacteria, viruses, parasites, antigens, specific antibodies,
nucleic acid sequences and the like. Furthermore, such assay
procedures can include the steps without limitation of labeling,
amplifying and detecting such target molecules or particles. In
particular, detection procedures including, but not limited to
polymerase chain reaction (PCR), real-time PCR, ligase chain
reaction (LCR), strand displacement amplification (SDA), and
nucleic acid sequence based amplification (NASBA).
[0043] The present invention provides a system for automated
performance of bio-chemical assays that includes a disposable
fluidic cartridge (1) comprising of at least one reaction chamber
(3) connected to a network of fluidic channels (2) with at least
one inlet and one outlet channel. The said channels end at the down
side of the fluidic cartridge with at least two connecting pores
(4), (4').
[0044] This simplicity is first translated into the fluidic
cartridge design that can be composed from a plastic molded parts
comprising, as shown in FIG. 1, from at least one layer 1(a) with
fluidic structures and a sealing layer 1(b). In a preferred
realization of the invention, the closing down layer is composed
from a flexible polymeric material. With this respect, the flexible
closing layer is preferably composed from an elastomer and more
preferably from a thermoplastic elastomer. For the fluidic
cartridge working, the said flexible polymeric material forming the
lower side of the fluidic cartridge will further serve as an
interface (16) to seal the connection of the fluidic cartridge
pores (4'), (4') with the respective pores of the disposable vessel
(7) and fluidic manifold (13).
[0045] The structured layer of the fluidic cartridge 1(a) comprises
the fluidic network and the reaction chamber according to the
invention. The layout of a preferred realization of such structured
layer is shown in FIG. 2. The structured layer is composed from a
fluidic channel network (2) connected to a reaction chamber (3).
This connection is operated through an inlet and outlet channel
(25in) and (25out) respectively. The fluidic channel network ends
at the down sides of the chips in connection pores (4).
[0046] Operation of the fluidic cartridge as shown in FIG. 2
consists of aspirating the sample within the sample vessel (5)
connected to the pore (4'a), into the reaction chamber (3) through
the inlet channel (25in). This is achieved using an active pump
connected to the outlet channel (25out) through a connection pore
(4b). After being processed in the reaction chamber, the sample can
be transferred to a waste container (15) connected to the manifold
through a connection pore (13') as shown in FIG. 1.
[0047] The fluidic cartridge according to another preferred
embodiment of the invention further comprises a second sample
connection pore (4'b) connected to the reaction chamber (3) through
a second outlet channel (26out). The second outlet channel (26out)
forms a diverging branch of the first inlet channel (25in). In
operating conditions, the connection pore (4'b) is directly
connected to a recovery sample vessel (5') (as shown in FIG. 3)
that will be used to recover the reaction product(s) from the
reaction chamber through the outlet channel (26out). This can be
assured by pushing air using a second active pump element connected
to a second inlet channel (26in) of the reaction chamber through a
second connection pore (4a) and wherein the said second inlet
channel (26in) forms a diverging branch of the first outlet channel
(26out). As a typical example, but not limited to, the recovered
material can comprise purified bio-molecules as nucleic acids or
proteins.
[0048] The fluidic cartridge according to the invention further
comprises an injection pore (4'c) through which different generic
reagents, like but not limited to washing, lysis, binding or
detection reagents, can be transferred to the reaction chamber by
the aspiration pump connected to the pore (4b).
[0049] According to a preferred embodiment of the invention, the
reaction chamber contains a solid support with an active surface to
attach or capture target molecules or particles carried by the
sample. The said solid support includes but not limited to
particles (preferably magnetic) or porous matrix.
[0050] From the preceded, the invention discloses a fluidic
cartridge for purifying target biomolecules from a sample that
comprises: [0051] (a) at least one structure top layer containing:
[0052] i. a reaction chamber (3) with a solid support that is
designed to capture the said target biomolecules, [0053] ii. a
first inlet (25in) and outlet (25out) channel that are in fluid
communication with the said reaction chamber and that will be used
to bring the sample into and out of the reaction chamber, [0054]
iii. a second inlet (26in) and outlet (26out) channel connected to
the said reaction chamber and that will be used for eluting the
purified biomolecules [0055] iv. wherein the second inlet and
outlet channels are diverging branches of the first inlet and
outlet channels, [0056] (b) a closing down layer composed from a
flexible material and comprising connection pores associated to the
ends of the said inlet and outlet channels.
[0057] In a preferred embodiment, the closing down layer of the
fluidic cartridge further can be used as an elastic membrane (27)
that can be deformed using an external actuation to seal one
specific channel and thereby prevent the flow through the said
channel. This is particularly important in operating the chip,
where by using this valve mechanism it allows for instance to seal
the recovery channel (26out) or the sample inlet channel (25in)
during the sample aspiration out of the sample tube (5) into the
reaction chamber (3) or the eluate sample recovery from this
reaction chamber into the recovery tube (5') respectively.
[0058] Accordingly, the present invention further provides a system
for automated conducting of bio-chemical assays that includes at
least one disposable vessel (5) comprising of a connection tube
(22) immersed in a sample container (6) and ending at the cap of
the vessel with an external connection pore (7a).
[0059] Rather than to be directly integrated into the cartridge,
the simplicity of the systems consists of the use a disposable
sample vessel container (5) composed from a traditional laboratory
sample collection tubes (25) with a pierceable cap (23). To assure
the connectivity with the fluidic cartridge a syringe adapter (22)
is inserted into the tube. It will be obvious for a person having
ordinary skills in the art that the sample containing vessel can be
alternatively manufactured as one piece with the connection tube
already integrated into the vessel core.
[0060] In a preferred embodiment, such tubes will be preferably
used for containing the starting sample that will be assayed.
[0061] Preferably also, the collection tube according to the
invention can be used as a tube (5') for recovering specific
molecules as for instance purified bio-molecules or particles after
their separation in the fluidic cartridge. In this particular
context, the recovery tube can be also used as an intermediary tube
that can contain a specific liquid that will further react with the
sample after being eluted out of the reaction chamber. Such
reaction can include but not limited to, a buffering re-adaptation
reaction, enzymatic reaction, target amplification reaction and a
detection reaction. Furthermore, after reacting within the said
recovery tube, the resulting reactant can further be pumped out
into the fluidic cartridge for further assay processing steps.
[0062] In a preferred embodiment according to the invention, the
automated system assembly for conducting bio-chemical assays can
further include at least one disposable vessel (5') ending at the
cap of the vessel with an external connection pore (7b) and that
can be used for recovering reactants as purified bio-molecules out
of the fluidic cartridge. The difference of this tube as compared
with the tube according to the preceding embodiments is that the
reactant cannot be further pumped out into the fluidic cartridge
for further assay processing steps.
[0063] Accordingly, the present invention further provides a system
for the automated performance of bio-chemical assays that includes
a fluidic manifold (8) that is part to the bulk system (12)
comprising of a fluidic channel network (9) and active fluidic
components (10), (11). The said channel network ends at the top
side of the fluidic manifold with at least one connecting pore (13)
and wherein the first and the second pore (4'), (4) of the fluidic
cartridge are interfaced by direct contact with the sample (7) and
the manifold (13) container pores respectively.
[0064] In a preferred embodiment, the fluidic manifold (8)
comprises of a fluidic network channels (9) and of active fluidic
elements like valves (11) and pumps mounted on the said manifold
(10). The fluidic manifold according to the invention is further
characterized by the fact that it is part of the bulk system (12).
Fluidic connectivity of the fluidic manifold (8) is assured through
connection pores (13) disposed on the top side of the manifold. The
said connection pores (13) are designed in a way to be aligned with
corresponding connection pores (4) positioned on the down side of
the fluidic cartridge. With this respect and as shown in FIG. 5,
the manifold pores (13a), (13b) and (13c) will be respectively
connected to the fluidic cartridge (4a), (4b) and (4c). As already
described, the connections (13b)-(4b) will assure the functions of
aspirating the sample from the vessel (5) to the reaction chamber
(3) within the fluidic cartridge (1) while the pore connectivity
(13a)-(4a) will be used to recover the reaction products to the
vessel (5') by injecting air to pouch the eluate out of the
reaction chamber into the recovery tube. Additionally, the
connection (13c)-(4c) can be used to inject reagents (like but not
limited to washing, lysis, binding, enzymes, buffers . . . ) into
the fluidic cartridge and therefore the reaction chamber by the
aspiration pump connected to the pore (13b)-(4b). As shown in FIG.
3, the reagents are preferably provided in disposable containers
(15) that are in fluidic connection with the manifold.
[0065] In a preferred embodiment and as shown in FIG. 3, the
reagent disposable containers (15) are first introduced in a
locking part (18) composed from a moving part comprising connecting
syringes (20) with a supporting fluidic part (19). The supporting
fluidic part (19) comprises fluid transfer channels associated with
each syringe. Each transfer channel of the fluidic part is
connected to the fluidic manifold (8) by classical tubing and
fittings (not shown). By that, a series of solenoid valves (11)
mounted on the manifold will be used to select one specific reagent
that will be transferred from the respective reagent container (15)
to the manifold pore (13c) and thereby be injected into the fluidic
cartridge (1).
[0066] In preferred embodiment, the manifold can not only used as a
support to transfer the sample(s) and the reagents into and from
the fluidic cartridge but also to inject reagents or recover eluate
into the samples vessels (5)-(5'). For instance, a reagent can be
injected through the pore connectivity (13c)-(4c) from a reagent
container (15) into the fluidic cartridge to be thereafter
transferred in a sample vessel (5) or (5').
[0067] As described by the different embodiments, the disclosed
fluidic assembly permit to perform extremely complex assay
procedures and reagents combinations in a very simple overall
system design.
[0068] For more precise positioning and control of the different
liquids in the reaction cartridge, in a preferred embodiment of the
invention, the fluidic manifold further integrates optical sensors
(29), (29') as shown in FIG. 5. As illustrated in FIG. 7 the
optical sensors will be placed on both side of the reaction chamber
(3) facing windows of detection channels (28) and (28'). The role
of the optical sensor (29) is to detect the presence of the liquids
in the detection windows (28). For that, a standard proximity
sensor can be used and that comprises of a photodiode and a laser
diode (LED). As shown in FIG. 8, the presence of the liquid in the
detection window (FIG. 8b) channel (28) leads to a change in the
reflection of the light emitted by the LED when compared with the
case liquid absence ((FIG. 8a). This change of the light reflection
induces a drop of the signal detected by the photodiode in case the
liquid is present in the detection window. This sensing process can
be used to determine the position of the liquid upstream and
downstream of the reaction chamber, which is very important of the
control and management of the assay procedures.
[0069] The liquid position according to the invention is
particularly required in assays based on magnetic particles where
the magnetic particles handling is particularly sensitive to the
presence or absence of the liquid. The magnetic particles handling
are preferably handled according to the device and the method
disclosed in the international patents applications WO2008/010111
and WO2008/007270, incorporated herein as a reference.
Specifically, this method utilizes at least one couple of
electromagnetic poles having magnetic field sequences having
polarity and intensity that vary in time, the role of which is to
effectively break or control the particle aggregates and to
maintain the particles in suspension as a fog of particles in
relative dynamic motion; and then combining the magnetic fields
from different magnetic poles in a sequence to induce displacement
of the fog of particles across the reaction chamber whereby the fog
of particles occupies substantially the whole reaction chamber
volume. In fact, when the magnetic particles are homogenously mixed
within the reaction chamber it is important to assure that the
chamber is fully filled with liquid to avoid an inadequate filling
during the process.
[0070] From the preceded, the invention discloses a fluidic
assembly for conducting bioassays which comprises a fluidic
cartridge with a reaction chamber and at least one optical sensor
positioned on one side of the said reaction chamber. In the
preferred embodiment, the optical sensor is a proximity planar
sensor comprising of an emitting Laser diode (LED) and a detection
photodiode. Accordingly, the reaction chamber preferably further
comprises magnetic particles that serve as a solid support for
performing the said bioassays.
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