U.S. patent application number 10/741628 was filed with the patent office on 2004-12-30 for automated sample processing apparatus and a method of automated treating of samples and use of such apparatus.
This patent application is currently assigned to DakoCytomation Denmark A/S. Invention is credited to Favuzzi, John, Key, Marc, Lathrop, Bob, Rasmussen, Ole F., Winther, Lars.
Application Number | 20040266015 10/741628 |
Document ID | / |
Family ID | 32682266 |
Filed Date | 2004-12-30 |
United States Patent
Application |
20040266015 |
Kind Code |
A1 |
Favuzzi, John ; et
al. |
December 30, 2004 |
Automated sample processing apparatus and a method of automated
treating of samples and use of such apparatus
Abstract
An automated staining apparatus and a method for treating
samples arranged on a carrier is disclosed, in which portions of
two or more selected reagents from a first station containing the
reagents are collected individually by a robot arm handling a probe
and dispensed in a mixing station of the apparatus. After mixing
the reagents in the mixing station, e.g. by shaking or rotating the
mixing cup containing the reagents, the reagent mixture is applied
to selected samples by the robot arm and a probe.
Inventors: |
Favuzzi, John; (Santa
Barbara, CA) ; Key, Marc; (Ojai, CA) ;
Winther, Lars; (Smoerum, DK) ; Rasmussen, Ole F.;
(Malov, DK) ; Lathrop, Bob; (San Jose,
CA) |
Correspondence
Address: |
SANTANGELO LAW OFFICES, P.C.
125 SOUTH HOWES, THIRD FLOOR
FORT COLLINS
CO
80521
US
|
Assignee: |
DakoCytomation Denmark A/S
Glostrup
DK
|
Family ID: |
32682266 |
Appl. No.: |
10/741628 |
Filed: |
December 19, 2003 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
60435601 |
Dec 20, 2002 |
|
|
|
Current U.S.
Class: |
436/48 ;
422/65 |
Current CPC
Class: |
Y10T 436/112499
20150115; Y10T 436/2575 20150115; Y02A 90/22 20180101; Y10T
436/114998 20150115; Y10T 436/113332 20150115; Y10S 901/41
20130101; G01N 2035/0091 20130101; G01N 2035/00366 20130101; G01N
35/00732 20130101; G01N 1/312 20130101; G01N 35/00871 20130101;
Y02A 90/26 20180101; G01N 35/00712 20130101; G01N 2035/009
20130101; Y10T 436/11 20150115; G16H 50/20 20180101; Y10T 436/25
20150115; G01N 2035/00376 20130101; G01N 2035/00881 20130101; G01N
1/31 20130101; G01N 35/0092 20130101; G01N 35/0099 20130101; G01N
2035/00326 20130101; G01N 1/30 20130101; G01N 2035/00891 20130101;
Y02A 90/10 20180101; G01N 2035/00089 20130101; Y10S 901/02
20130101; G01N 2035/00306 20130101; G01N 2035/00039 20130101; G01N
2035/00435 20130101; Y10T 436/114165 20150115 |
Class at
Publication: |
436/048 ;
422/065 |
International
Class: |
G01N 035/02 |
Claims
What is claimed is:
1. A sample processing apparatus for treating samples arranged on a
carrier comprising: a vial station for containing at least two
reagent vials, a carrier station arranged for intermediate storage
of a plurality of carriers, a probe drive adapted to move a probe,
wherein the probe drive is arranged to aspirate a portion of
reagent from a selected reagent vial of the vial station with a
probe and to apply reagent to a selected carrier of the carrier
station, a reagent mixer having a mixing cup adapted to receive two
or more reagents and mix element adapted to mix the reagents in the
mixing cup, wherein said reagent mixer is adapted to dispense the
reagent mixture from the mixing cup to a selected carrier of the
carrier station.
2. A sample processing apparatus according to claim 1, wherein the
probe drive is arranged to apply the reagent mixture from the
mixing cup to selected carrier of the carrier station.
3. A sample processing apparatus according to claim 1 or 2, wherein
the probe drive is arranged to aspirate portions of reagents from
at least two selected reagent vials of the vial station and
dispense said portions of reagents into the mixing cup.
4. A sample processing apparatus according to any of claims 1 to 3,
wherein the reagent mixer further comprises cup drive arranged for
cyclic movement of the mixing cup so as to mix reagents contained
in the mixing cup.
5. A sample processing apparatus according to claim 4, wherein the
cyclic movement is a rotation of the mixing cup and wherein the
rotation is an intermittent rotation in a clockwise and in an
anticlockwise direction.
6. A sample processing apparatus according to claim 5, wherein the
rotation takes place about a substantially vertical axis.
7. A sample processing apparatus according to claim 4, wherein the
movement is a combination of rotations with variable speed, various
speeds, stops and change of direction, wherein the rotations is
controlled by control element according to the sample processing
protocol for the sample(s) processed in the apparatus.
8. A sample processing apparatus according to claim 3, wherein the
mixing cup has inner walls extending upwardly and outwardly from a
bottom part of the mixing cup, and the cup drive is capable of
rotating the mixing cup at an angular speed sufficient to fling
reagents contained therein out of the mixing cup.
9. A sample processing apparatus according to claim 8, wherein the
reagent mixer comprises a waste reagent collecting chamber having a
sidewall part laterally surrounding the mixing cup, and wherein
waste reagent collecting chamber is arranged to collect reagents
flung out of the mixing cup by rotation of the cup.
10. A sample processing apparatus according to claim 3, wherein the
reagent mixer further comprises a mixing cup holder adapted to
receive and support the mixing cup in a releasable manner.
11. A method of fully automated treating of samples arranged on a
carrier of a sample processing apparatus controlled by control
element, wherein the method comprises the steps of: arranging a
plurality of carriers intermediately in a carrier station, each
carrier having a sample arranged thereon, dispensing a portion of a
first reagent selected from a plurality of reagents to a mixing
cup, dispensing a portion of a second reagent selected from a
plurality of reagents to the mixing cup, mixing the reagents in the
mixing cup, moving the probe to the mixing cup with a probe drive,
aspirating a portion of the mixed reagents from the mixing cup with
the probe, moving the probe to a selected one of said carriers, and
dispensing the mixed reagents to the selected carrier, so as to
perform a treatment of the sample arranged on the selected
carrier.
12. A method according to claim 1 1, wherein the steps of dispense
portions of the first and the second reagents comprises the steps
of: moving a probe to a first, selected reagent vial with the probe
drive, aspirating a portion of the first reagent from the first
reagent vial with the probe, moving the probe to the mixing cup
with the probe drive, dispensing the portion of the first reagent
to the mixing cup, moving a probe to a second, selected reagent
vial with the probe drive, aspirating a portion of the second
reagent from the second reagent vial with the probe, moving the
probe to the mixing cup with the probe drive, and dispensing the
portion of the second reagent to the mixing cup.
13. A method according to claim 11 or 12, further comprising the
step of controlling the rotation pattern according to the sample
processing protocol thereby optimising the quality the mixing in
the mixer cup in respect to the reagents present in the mixer
cup.
14. A method according to claim 11 or 12, further comprising the
step of adding a protective gas covering the reagent mixture in the
mixer cup.
15. A method according to claim 14, wherein the protective gas is
selected from the the group consisting of noble gasses, nitrogen,
and carbon dioxide.
Description
[0001] This application claims the benefit of U.S. Provisional
Application No. 60/435,601, filed Dec. 20, 2002, hereby
incorporated by reference.
FIELD OF INVENTION
[0002] This application relates to the field of sample processing
systems and methods of processing samples. The present invention
may be directed to the automated processing, treatment, or even
staining of samples arranged on carriers, such as slides, and in
some embodiments, directed to the continuous or batch processing of
samples and carriers, as well as washing elements of a sampling
system. Embodiments may further relate to control systems for
sample processing and data acquisition, data maintenance, and data
retrieval for sample processing. Applications to which the present
invention may especially relate include immunohistochemistry,
in-situ hybridization, fluorescent in-situ hybridization, special
staining, and cytology, as well as potentially other chemical and
biological applications.
[0003] More specifically the present invention relates to an
automated sample processing apparatus for treating samples arranged
on carrier means, such as microscope slides or sample tubes,
located at defined positions close to or in the apparatus by
aspirating a portion of selected reagent from a station containing
a plurality of reagents and thereafter dispense the reagent to a
sample, e.g. a tissue, organic cells, bacteria etc., arranged on
the carrier means. The invention facilitates that two or more
reagents are mixed and the mixture applied to a sample.
[0004] The invention also relates to a method of automated treating
of samples by mixing reagents and dispensing the mixture to the
sample.
[0005] A sample is in this application to be understood as a
biological sample such as histological samples, e.g. tissue and
cell specimens, including cell lines, proteins and synthetic
peptides, tissues, cell preparations, blood, bodily fluids, bone
marrow, cytology specimens, blood smears, thin-layer preparations,
and micro arrays, and specifically biological samples on microscope
slides.
BACKGROUND OF INVENTION
[0006] Sample processing in immunohistochemical (IHC) applications
and in other chemical and biological analyses may require one or a
number of various processing sequences or protocols as part of an
analysis of one or more samples. The sample processing sequences or
protocols may be defined by the individual or organization
requesting an analysis, such as a pathologist or histologist of a
hospital, and may be further defined by the dictates of a
particular analysis to be performed.
[0007] In preparation for sample analysis, a biological sample may
be acquired by known sample acquisition techniques and may
comprise, for example in IHC applications, tissues generally or
even in some applications one or a plurality of isolated cells,
such as in microarray samples, and may be presented on a sample
carrier such as a microscope slide. Furthermore, the sample may be
presented on the carrier variously and potentially in some form of
preservation. As one example, a sample such as a layer or slice of
skin may be preserved in formaldehyde and presented on a carrier
with one or more paraffin or other chemical layers infiltrating the
sample.
[0008] Immunologic and histological applications, for example, may
require processing sequences or protocols that comprise steps such
as deparaffinization, target retrieval, and staining, especially
for in-situ hybridization (ISH) techniques. Previously, in some
applications, these steps may have been performed manually,
potentially creating a time-intensive protocol and necessitating
personnel to be actively involved in the sample processing.
Attempts have been made to automate sample processing to address
the need for expedient sample processing and a less manually
burdensome operation. However, such previous efforts may have not
fully addressed the needs for an automated sample processing
system. Previous efforts to automate sample processing may be
deficient in several aspects that prevent more robust automated
sample processing, such as: the lack of sufficient computer control
and monitoring of sample processing; the lack of information
sharing for processing protocol and processing status, especially
for individual samples; the lack of diagnostic capabilities; and
the lack of real-time or adaptive capabilities for multiple sample
batch processing.
[0009] Past efforts at automated sample processing for samples
presented on carriers such as slides, such as U.S. Pat. No.
6,352,861 to Ventana Medical Systems, Inc. and U.S. Pat. No.
5,839,091 to LabVision Corporation, have not afforded the various
advantages and other
[0010] Sample processing apparatuses for staining and treating
samples by means of probes normally comprises a first station for
containing one or more reagent vials; a second station for mounting
slides, a probe arranged to aspirate a portion of reagent from a
selected reagent vial and dispensing the reagent to a slide on
which the sample is arranged and a drive means for moving the probe
between the various stations.
[0011] U.S. Pat. No. 5,948,359 discloses an apparatus of the above
mentioned type, wherein the first station comprises a vial holder
for holding 40 or more vials in order to provide a wide range of
different reagents adapted for different staining purposes, and
thereby the possibility of automatically staining a large number of
slides requiring different staining processes. In practise it is
very important that the apparatus facilitates that many different
staining processes can be performed at the same time in the
apparatus, because this avoids the necessity of batching samples
requiring the same procedure or other treatment with reagents, and
processing each batch individually.
[0012] U.S. Pat. No. 5,723,092 discloses a sample dilution well for
an immunoassay analyser in which a sample and a diluent is mixed by
rotating the sample dilution well. Samples such as plasma or urine
are loaded into the immunoassay analyser in a number of sample
containing tubes, and a probe is arranged for collecting a portion
of the sample in a selected sample carrying tube and transferring
the sample to the dilution well, adding a portion of diluent, such
as water to the sample portion and mixing the two by rotating the
dilution well, after which a portion of the diluted sample is
aspirated from the dilution well for examination.
[0013] The mixing of reagents is known from U.S. Pat. No. 5,425,918
disclosing a staining apparatus for treating samples arranged on
carrier means, comprising a vial station for containing at least
two reagent vials, a carrier means station arranged for
intermediate storage of a plurality of carrier means, probe drive
means arranged for moving a probe, wherein the probe drive means is
arranged to aspirate a portion of reagent from a selected reagent
vial of the vial station by means of a probe and to apply reagent
to a selected carrier means of the carrier means station. In this
document it is suggested how to solved the problem regarding agents
requiring mixing just prior to use due to short active lifetimes
after mixing. The agents may be mixed by use of the probe and probe
drive means drawing up one agent from its storage vial and
dispensing it into the storage vial of the second agent. The mixed
solution is then drawn up and dispensed onto the slides.
[0014] Mixing wells for mixing samples and diluents are also
disclosed by U.S. Pat. No. 4,323,537, in which an analysis system
is described having mixing cuvettes for mixing sample and diluents
by rotation and a displaceable probe for collecting the raw sample
from a sample container and deliver it to the mixing cuvette, and
for collecting the diluted sample from the cuvette after mixing and
deliver it to a plurality of analysis stations for further,
automated analysis.
[0015] Methods for mixing reagents and liquids are well known--but
several aspects are important in the context of IHC and ISH
instruments wherein the mixing in some processes can be
cumbersome.
[0016] The staining procedure is laborious and uses many different
reagents. The staining protocol may include the following steps:
deparaffination, washing, antigen retrieval, endogenous biotin or
enzyme blocking, incubation with immunological reagents, molecular
probes, secondary visualization reagents and various chromogen
reagents, washing steps and counterstaining.
[0017] An on-board mixing device should be able to mix a multiple
of reagents and mixtures. Non-limiting examples include:
[0018] Dilution of chromogens concentrates, mixing and dilution of
two, three or four component enzyme chromogen reagents, dilution of
buffer concentrates, dilution of immunological reagents with
dilution buffer, dilution of visualization reagents with dilution
buffers, mixing several visualization reagents with dilution buffer
or dilution of enzyme blocking reagents, dilution of biotin
blocking reagents or mixing and dilution of counterstaining
reagents.
[0019] During the mixing several problems may arise. Some problems
arise due to the complex use of reagents in the staining procedure.
Below a few non-limiting examples are listed in more detail:
[0020] The chromogen reagents (e.g. DAB, AEC, fast red etc) often
comes as concentrated reagents in organic or high viscosity
solutions and needs to be diluted prior to being applied to the
sample.
[0021] Chromogens like the Fast Red alkaline phosphatase chromogen
are made ready for use by mixing and dilution of two or three
reagents, which are very different in nature with regard to salt
content, viscosity and density. Furthermore, the resulting mixtures
are unstable over time and need to be used within a short time.
[0022] Some chromogens suitable for e.g. horseradish peroxidase,
like DAB and AEC, are easily oxidized when exposed to air during
e.g. vigorous mixing or dilution.
[0023] The enzyme chromogens and counterstain reagents like e.g.
hematoxylin, are semi oxidized and can contain precipitates and
solids. By further oxidation or slight change in pH, the reagents
can further precipitate.
[0024] Antibody and enzyme containing reagents often contain
stabilizing proteins and or detergents, which causes the solution
to foam when being shaken or stirred. Many proteins cannot easily
tolerate to be exposed to the hydrophobic air in foam.
[0025] Wash buffers can contain detergents, which can foam when
shaken or stirred. The foam can spread to other compartments of the
instrument in an unwanted and unpredictable way.
[0026] Mixing of some reagents like e.g. the HRP chromogens and
peroxide reagents can result in the formation of small bobbles.
These can generate foam or bubbling on the surface of the
mixture.
[0027] Spill over/carry over must be avoided. The staining process
is characterized by using many, complex and very different reagents
and buffers and in many different dilution ratios and mixtures.
Some of the reagents or buffers are incompatible with each other's.
In the event of cross contamination due to e.g. carry over, the
reagents may be ruined within seconds or solids can precipitate,
making the staining unsuccessful. For example, enzyme containing
reagents can not be mixed with the corresponding chromogens, or
high salt concentrates may not be mixed with e.g. proteins
containing mixtures, or organic solvents can not be mixed with
protein containing mixtures, or highly pH buffered wash buffers can
not be mixed with low buffered mixtures without significantly
altering the properties of the reagents. Accordingly the cleansing
and washing of the mixing device need to be very efficient.
[0028] As the procedures are very complex, and the instrument uses
many different protocols, one cannot predict the result of reagent
carry-over or unplanned mixing of reagents. Consequently, the
mixing device should ideally be very efficient and be designed for
a variety of reagent mixing protocols and sequences.
[0029] Build-up of small fouling layers on the surfaces will fast
cause problems, as the typical staining protocol calls for many
mixing and dilution steps. Consequently, the mixing device should
have a minimum of surface area and very smooth surfaces.
[0030] Furthermore, the mixing device should ideally be able to mix
very different volumes of reagents in both small and large volumes
ratios. In other words, the degree of dilution and mixing ratios of
reagents may vary from small to high ratios.
[0031] In summary, the mixing device should ideally allow:
[0032] Mixing of small and large volumes
[0033] Mixing reagents with different viscosities and densities
[0034] Mixing of immiscible or nearly immiscible reagents
[0035] No fouling of mixing rods or similar due to precipitated
material
[0036] Allow easy escape of formed gasses during mixing
[0037] Prevent foaming of e.g. detergent or protein containing
reagents
[0038] Low build-up of debris or fouling on the device surfaces
[0039] Easy emptying and washing--regardless of volumes
[0040] Very low reagent carry-over
[0041] No present mixing system truly fulfils the above-mentioned
properties. On the slide mixing does not allow for very large
ratios of dilution. Nor does it allow for efficient mixing of
reagents with very different densities or viscosity.
[0042] The object of the present invention is to improve the known
apparatuses for sample processing as well as the method for
automatic sample processing by facilitating a wider range of
available processes of treatment, so as to increase the number of
different staining and/or treatment processes that may be performed
automatically, alternatively or additionally to provide an
increased quality of some specific staining processes.
SUMMARY OF INVENTION
[0043] By providing a staining apparatus having an automated mixer
integrated therein, staining or treatment processes requiring a
mixture of unmixable reagents, such as a water based and an oil
based reagent, or insoluble reagents, can be performed
automatically instead of requiring human interaction or manual
performance of some process steps in a much more automated process,
and the quality of the staining process may be improved as a
desired degree of mixing of reagents may be provided or an optimal
application time window for a deteriorating mixture may be
reached.
[0044] This is achieved by the sample processing apparatus
according to the present invention, comprising a reagent mixer
having
[0045] a mixing cup for receiving or more broadly adapted to
receive two or more reagents and mixing means for mixing or more
broadly a mix element adapted to mix the reagents in the mixing
cup, and
[0046] means for dispensing the reagent mixture from the mixing cup
to a selected carrier means.
[0047] An apparatus according to the invention may comprise an
automated sample processing system comprising a plurality of
drawers, a plurality of sample carrier retainment assemblies each
removably configured with one of the drawers, and an adaptive
sample processing control system to which the drawers and the
sample carrier retainment assemblies are responsive. The sample
carrier retainment assemblies may comprise slide retainment
assembly and may be removably configured with the drawers to
provide sample processing with the drawers of the system. The
adaptive sample processing control system may automate the sample
processing system such that one or more batches of samples may be
processed according to one or more protocols, potentially indicated
by information on the slides that may be automatically identified
by the adaptive sample processing control system. Sample processing
may comprise one or more sampling protocols and steps, such as
deparaffinization, target retrieval, and staining.
[0048] The term staining is used for the end product of the
process, by which certain parts of the sample may be stained, i.e.
have obtain a different colour, either in the optic range or in
another electromagnetic range, such as ultra violet, or the
staining may be an detectable, preferably automatically detectable,
change in properties, such as fluorescent properties, magnetic
properties, electrical properties or radioactive properties. To
obtain the staining, the sample normally have to undergo a series
of treatment steps, such as--but not limited to--washing, binding
of reagents to the specific parts of the sample, activation of the
reagents, etc. and each treatment step may include a plurality of
individual treatments.
[0049] In some staining processes, it is required for one or more
treatments to use a mixture of reagents prepared from two or more
separate reagents which may be somewhat incompatible e.g.
unmixable, such as a water based and an oil based reagent, or
insoluble, and therefore requires that the two or more reagents are
manually prepared and introduced into a reagent vial shortly before
starting the staining process in order to obtain the best possible
staining result for the selected examination purposes. For other
processes, different staining process steps require a mixture of
the same two reagents but in different dissolution ratios. Some
process step may require a mixture of two or more reagents that,
when mixed, becomes unstable, e.g. has a limited time window of
usability because internal chemical processes deteriorates the
mixture. By providing a sample processing apparatus having an
automated mixer integrated therein, these types of staining
processes can be performed automatically instead of requiring human
interaction or manual performance of some process steps, thereby
achieving a much more automated process, and the quality of the
staining process may be improved as a desired degree of mixing of
reagents may be provided as well as an optimal application time
window for a deteriorating mixture may be reached.
[0050] Thus, the present invention relates to a sample processing
apparatus for treating samples arranged on carrier means or
carrier, comprising
[0051] a vial station for containing at least two reagent vials, a
carrier means station or more broadly a carrier station arranged
for intermediate storage of a plurality of carrier means or
carriers,
[0052] probe drive means, or more broadly a probe drive arranged
for moving or more broadly adapted to move a probe,
[0053] wherein the probe drive means is arranged to aspirate a
portion of reagent from a selected reagent vial of the vial station
by means of or even with a probe and to apply reagent to a selected
carrier means of the carrier means station,
[0054] wherein the apparatus comprises a mixing station with a
reagent mixer having a mixing cup for receiving or adapted to
receive two or more reagents and mixing
[0055] means for mixing or more broadly a mixer element adapted to
mix the reagents in the mixing cup, and
[0056] means for dispensing the reagent mixture from the mixing cup
to a selected carrier means of the carrier means station.
[0057] The vial station is a collection of a plurality of vials, at
least two, but often 20-60 vials or more, which may or may not be
physically arranged in close proximity to each other. The term
station does not indicate that the vials must be located within
one, confined area; rather it indicates the existence of a
plurality of vials. The probe drive means or even probe drive may
be a robot arm with two or three degrees of freedom, such as an
articulated arm or one track or a set of perpendicular tracks along
which a probe retainer of the probe drive means may be displaced,
wherein the probe retainer may be moved in a direction normal to
the track or tracks. The skilled person may readily design other
types of probe drive means, e.g. combinations of the above
described. The carrier means or even carrier may be provided to the
apparatus in a two-dimensional array, e.g. constituted by
individual rows of carrier means as discussed in the example below,
or the carrier means may be provided in any manner known in the
art, e.g. arranged in a carrousel or as a row of carrier means. The
carrier means may also be arranged movably with respect to the
probe drive means, such as in an endless row that is advanced
automatically past the operating area of the probe drive means or
as a two-dimensional array that may be moved in a direction
perpendicular to a travel direction of the probe drive means, so
that the probe may reach any carrier means by the combined movement
of the probe and the array.
[0058] The carrier means are preferably arranged in groups or
series on trays or the like, so that a plurality of carrier means
may be removed from or inserted into the apparatus simultaneously,
and the apparatus preferably also comprises means for performing
the intermediate storage of the carrier means with samples thereon
and the removal of the carrier means from the apparatus
automatically.
[0059] The operation of the sample processing apparatus will
generally be controlled by means of control means, typically a
computer having a central processing unit and one or more memory
unit associated therewith, means for controlling or more broadly a
control element the various operations of the apparatus by
controlling step motors, solenoids, valves and/or other drive or
control parts of the apparatus. The control means may have one or
more data communication ports for enabling data communication with
external computers by wire or wireless. The control means does not
have to be physically arranged within the apparatus itself but may
be a computer external to the sample processing apparatus and
connected to the apparatus via a data transmission port
thereof.
[0060] It is advantageous that the probe drive means is arranged to
apply the reagent mixture from the mixing cup to selected carrier
means. The mixed reagents from the mixing cup may be applied to the
samples by separate means, such as a separate probe and probe drive
means. In an alternative embodiment the carrier with the sample in
question may be moved to a location at an outlet from the mixing
cup. However it is presently preferred to use the probe drive means
for the task.
[0061] The mixing cup may receive the two or more reagents to be
mixed from a separate set of vials or other reagent sources, but it
is for rationalisation of the apparatus preferable that at least
some of the reagents to be mixed may come from the same vials as
are used for containing reagents to be applied to the samples. In a
preferred embodiment, the probe drive means is therefore arranged
to aspirate portions of reagents from at least two selected reagent
vials of the vial station and apply said portions of reagents to
the mixing cup.
[0062] The mixing means or more broady a mix element of the reagent
mixer may advantageously be constituted by cup drive means or more
broadly a cup drive arranged for cyclic movement of the mixing cup,
e.g. shaking or rotation in a horizontal or a vertical plane, so as
to mix reagents contained in the mixing cup. The mixing of the
reagent may be further improved by arranging mixing elements, such
as blades or edges within the cup and stationary with respect to
the cup. Other known mixer types may instead be preferred, such as
shaft-driven impellers or magnetically driven impellers.
[0063] The cyclic movement is preferably a rotation of the mixing
cup, advantageously about a substantially vertical axis. The
rotation is in a preferred embodiment an intermittent rotation in a
clockwise and in an anticlockwise direction. Preferably the
rotation speed may be varied through control means acting upon a
driving motor in such manner that the fluid content inside the
mixing cup is agitated, but at so low speed that substantially all
reagents remain in the mixer cup. Preferably the speed may be
controlled depending on the type of reagent in the mixer cup.
Preferably the speed is controlled according to the processing
protocol controlling the processing of the sample(s) to be
processed in the apparatus. Preferably the rotation pattern (speed
and direction) is controlled by controlling means or more broadly a
control element, such as a computer, according to the sample
processing protocol, thereby optimising the quality the mixing in
the mixer cup in respect to the reagents present in the mixer cup.
For the sake of good order, by sample processing protocol is meant
a sequence of processing steps defined for the actual sample
treated in the sample processing apparatus.
[0064] In order to provide an efficient and fully automated
cleansing of the mixing cup when changing from one reagent mixture
to the next, it is advantageous that the mixing cup has inner walls
extending upwardly and outwardly from a bottom part of the mixing
cup, and the cup drive means is capable of rotating the mixing cup
at an angular speed sufficient to fling all reagents contained
therein out of the mixing cup. Preferably the inner walls are
smooth and may terminate in an upper rim at the widest inner
diameter of the mixing cup, in which case the high-speed rotation,
preferably about a symmetry axis of the cup, will cause the waste
reagent or cleansing liquid to be flung out over this upper rim. In
an alternative embodiment of the mixer, the inner walls of the cup
extend upwards and inwardly above the level of the widest inner
diameter, at which one or more exit openings are provided in the
inner wall as outlets for the waste reagent or the cleansing
liquid. The top may be open for the probe to enter the mixing cup,
or it may be closed to facilitate that the cyclic movement of the
mixing cup for mixing the reagents may have a vertical component.
The probe may in this case enter the mixing cup through one of the
exit openings.
[0065] To collect the waste reagent or the cleansing liquid, it is
preferred that the reagent mixer comprises a waste reagent
collecting chamber having a sidewall part laterally surrounding the
mixing cup and arranged to collect reagents flung out of the mixing
cup by rotation of the cup.
[0066] In a further preferred embodiment, the reagent mixer further
comprises a mixing cup holder for receiving and supporting the
mixing cup in a releasable manner, so that different mixing cups
may be used for different reagent mixtures.
[0067] In yet further preferred embodiment, replacing means are
provided for replacing the probe that is moved by the probe drive
means with another probe, so as to avoid contamination of the
reagents in the various reagent vials by the use of more than one
probe. This may be combined with a probe washing station, in which
one probe is washed after it has been replaced at the probe drive
means so that it is clean and ready for repeated use without
risking contamination of the content of the reagent vials.
[0068] The present invention also relates to a method of fully
automated treating of samples arranged on carrier means by means of
a sample processing apparatus controlled by means of a control
means, wherein the method comprises the steps of:
[0069] arranging a plurality of carrier means intermediately in a
carrier means station, each carrier means having a sample arranged
thereon,
[0070] dispensing a portion of a first reagent selected from a
plurality of reagents to a mixing cup,
[0071] dispensing a portion of a second reagent selected from a
plurality of reagents to the mixing cup,
[0072] mixing the reagents in the mixing cup by means of mixing
means,
[0073] moving a probe to the mixing cup by means of a probe drive
means,
[0074] aspirating a portion of the mixed reagents from the mixing
cup by means of the probe,
[0075] moving the probe to a selected one of said carrier means,
and
[0076] dispensing the mixed reagents to the selected carrier
means,
[0077] so as to perform a treatment of the sample arranged on the
selected carrier means.
[0078] In a preferred embodiment, the steps of dispensing portions
of the first and the second reagents comprises the steps of
[0079] moving a probe to a first, selected reagent vial by means of
the probe drive means or even with a probe drive,
[0080] aspirating a portion of the first reagent from the first
reagent vial by means of the probe or even with a probe,
[0081] moving the probe to the mixing cup by means of the probe
drive means, dispensing the portion of the first reagent to the
mixing cup,
[0082] moving a probe to a second, selected reagent vial by means
of the probe drive means or even with a probe drive,
[0083] aspirating a portion of the second reagent from the second
reagent vial by means of the probe or even with a probe,
[0084] moving the probe to the mixing cup by means of the probe
drive means or even with a probe drive, and
[0085] dispensing the portion of the second reagent to the mixing
cup.
[0086] In a further preferred embodiment of the present invention,
the method further comprises the step of replacing the probe
arranged with the probe drive means with another probe between
handling two different reagents or between handling a reagent and a
reagent mixture.
[0087] The present invention further relates to the use of an
apparatus of the present invention as described above for
exercising the method of the present invention. More specifically
the invention relates to the use of the apparatus for IHC and ISH
and sample processing of the type using special stains as well as
batch-mode sample processing.
[0088] Support should be understood to exist for the following
aspect and embodiment of the invention: use of an apparatus
according to any of the apparatus claims for exercising the method
according to any of the method claims for immunohistochemistry and
in-situ hybridization. Also support should be understood to exist
for use of an apparatus according any of the apparatus claims for
exercising the method according to any of the method for batch-mode
sample processing.
BRIEF DESCRIPTION OF THE FIGURES
[0089] The invention will now be explained in further detail with
reference to the enclosed drawing, wherein
[0090] FIG. 1 is a perspective view of a sample processing
apparatus according to the invention,
[0091] FIG. 2 is a plan view of a sample processing apparatus
according to the invention,
[0092] FIG. 3 is a perspective view of a detail of the sample
processing apparatus according to FIG. 2,
[0093] FIG. 4 is a perspective view of a reagent mixer according to
the invention, and
[0094] FIG. 5 is a vertical cross-section of the reagent mixer
according to FIG. 4.
DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS OF THE INVENTION
[0095] As mentioned earlier, the present invention includes a
variety of aspects, which may be combined in different ways. The
following descriptions are provided to list elements and describe
some of the embodiments of the present invention. These elements
are listed with initial embodiments, however it should be
understood that they may be combined in any manner and in any
number to create additional embodiments. The variously described
examples and preferred embodiments should not be construed to limit
the present invention to only the explicitly described systems,
techniques, and applications. Further, this description should
further be understood to support and encompass descriptions and
claims of all the various embodiments, systems, techniques,
methods, devices, and applications with any number of the disclosed
elements, with each element alone, and also with any and all
various permutations and combinations of all elements in this or
any subsequent application.
[0096] The embodiment shown in the figures and described in details
below is only an example of an apparatus in accordance with the
present invention and is not limiting the wider scope of the
invention as described in the enclosed claims.
[0097] FIG. 1 shows one schematic embodiment of a sample processing
system 1 in accordance with the present invention. Cabinet sections
form outer portions of the system and serve to address general
structural considerations of the system (a top cabinet section is
not shown in FIG. 1). The sample processing system may comprise a
plurality of drawers used for the handling and processing of
samples and sample carriers such as slides, potentially microscope
slides. Other sample carriers may be accommodated consistent with
the present invention. Each drawer may be configured to accommodate
sample carrier retainment assemblies, such as slide retainment
assemblies, carrier racks, modules, or magazines.
[0098] One embodiment of a sample carrier retainment assembly may
comprise a slide retainment assembly. The slide retainment assembly
may comprise a slide rack, module, or magazines. Slide retainment
assembly is configured to accommodate a plurality of slides in at
least one configuration in corresponding sample carrier retention
devices. The sample carrier retainment assemblies, are utilized in
the processing of samples as further described below. It should be
further noted that the sample carrier retainment assembly can be
removably configured with the drawers, and may be stackable or
nested within other retainment assemblies.
[0099] One or more drawers may be provided to accommodate
processing materials such as reagent containers 3 for sample
processing, also further described below. A processing material
retainment assembly, such as a container rack may be utilized to
accommodate reagent containers or other processing materials within
each of drawers 6. Bottle inserts may be preferably configured with
the retainment assembly to ensure proper processing material
positioning within the processing material retainment assembly and
the drawer.
[0100] Multiple drawers allow for one or a plurality of sample
processing protocols to be performed by the system 1. Past efforts
at sample processing, as previously described, may have been
limited to processing sequences for an entire batch of carriers
within the system. The present invention, however, in part by
providing a plurality of drawers and carrier retainment assemblies,
may allow for multiple batch processing, including real-time or
adaptive capabilities for multiple batch processing, as further
described below.
[0101] The system may access, use and wash multiple probes or
syringes for pipetting or otherwise aspirating fluids required for
the staining of samples mounted or otherwise presented on slides.
To eliminate cross contamination a system with a single reusable
probe may wash the probe between each fluid applied. The task of
washing the probe can have a large impact on the throughput of the
overall system. The present invention may allow for multiple probes
to be available to the system for use. The system may continuously
have a clean, uncontaminated, sterilized, or an unused probe
available to use and sample processing is not impacted by the
required cleaning routine. The cleaning routine may be necessary to
eliminate the possible cross contamination of fluids and, in some
embodiments, may take up to about 1 minute to accomplish. The
cumulative impact of the cleaning routine on a series of processing
steps can add time to the throughput capabilities of the system.
The addition of multiple probes or syringes may eliminate this
impact and significantly decreases the time required to process the
samples.
[0102] Embodiments of the present invention comprise a mixing
station or reagent mixer 9 (best shown in FIGS. 4 and 5). The
system may mix component fluids, such as dyes, buffers, or other
processing materials, preferably on demand and as the processing
steps and protocols dictate. Fluids required during the processing
steps may sometimes need to be mixed with other fluids to create a
final activated fluid. However, the activity levels of these
mixtures can be time sensitive and may therefore only be effective
for a short period of time. The on demand mixing of fluids is
advantageous in that it allows the fluids to be mixed immediately
before being used. The syringe or probe 10, in preferred
embodiments, will aspirate fluids into and from the mixing station
9 to mix component fluids. A rinse may further be dispensed into
the mixing station to sterilize the station.
[0103] As an example a presently preferred embodiment of a sample
processing apparatus, also called a staining apparatus 1, according
to the invention is shown in FIGS. 1 and 2. The sample processing
apparatus 1 comprises a rectangular frame 4 surrounding a first
station 2 comprising an array of compartments wherein each
compartment a reagent vial 3 is placed, and a second station 5
wherein a number of separate racks 6 (which may be a drawer) is
placed, and where each rack comprises a number of microscope slides
7 mounted side by side in the rack 6. In the embodiment shown, each
rack may hold up to 8 slides, but the rack may be designed to hold
any suitable number of slides. With eight racks arranged side by
side, the shown embodiments may hold up to 64 slides 7 each having
a sample, e.g. a tissue mounted on the upper side of the slide, so
that reagent may be applied from above to the sample on each
slide.
[0104] A robot arm 20 in FIG. 1 for moving a probe 10 in X and Y
(as well as Z) direction as indicated by the arrows X and Y is
arranged above the frame 4 of the sample processing apparatus. The
robot arm may is therefore position the probe 10 above all reagent
vials 3 as well as above all the microscope slides 7, and may
further operate the probe 10 to aspirate portions of reagent
contained in any of the vials 3, to transfer the portion of reagent
and apply it to any of the slides 7 in order to provide a selected
staining or treatment of the sample on each slide 7. By use of
suitable control means e.g. a computer (not shown) having the
appropriate software and input data for the purpose, this staining
apparatus I is able to automatically staining or treating samples
requiring different staining or treatment reagents and
processes.
[0105] The same staining apparatus viewed from below the robot arm
is shown in FIG. 2, disclosing the probe 10 being manipulated by
the robot arm. The probe 10 is raised to an upper position (in a Z
direction where it is clear of the vials 3 underneath the probe 10,
but the robot comprises means (not shown) for lowering the probe 10
in order to dip the probe tip 211 into the content of a selected
reagent vial 3 and to aspirate a selected amount of reagent for the
selected staining or treatment process.
[0106] The staining apparatus 1 of the present embodiment further
comprises at least one probe washing station 8 and a reagent mixer
9, and the robot arm is furthermore arranged to transfer the probe
to the washing station 8 as well as to the reagent mixer 9. The
reagent mixer 9 will be described in detail below with reference to
FIGS. 4 and 5.
[0107] The apparatus comprises a reagent mixer 9 having a mixing
cup 213 wherein two or more selected reagents may be placed by
means of the robot arm and the probe 10. The reagent mixer 9
thereby provides on-board mixing of any reagents contained in the
reagent vials 3, and thereby more staining processes, e.g. staining
requiring the use of mixing of insoluble reagents, or reagents
which may only be effective a short time after mixing, are
facilitated to be performed automatically within the staining
apparatus without the requirement of human interaction.
[0108] The mixer 9 comprises a mixing cup 213 for receiving
reagents released from the probe 10 or 110. The mixing cup 213 is
placed into a holder 215 by means of a complementary snap fitting
means 216 and 217 arranged on the inside of the holder 215 and the
outside of the mixing cup 213, respectively. A motor 218 is
arranged for rotating the holder 215 and thereby the mixing cup
213, either intermittently clockwise and anticlockwise in order to
provide a mixing of reagents contained in the mixing cup, or by
spinning the holder 215 and thereby the mixing cup 213 in order to
fling out waste reagents or cleansing liquid from the mixing cup
213.
[0109] For the latter purpose, the mixing cup 213 is preferable
provide with sidewalls 220 extending upwardly and outwardly from
the bottom 219, e.g. forming a frusto-conical cavity, and the
mixing cup 213 has an upper rim 214 allowing the reagents to escape
from the mixing cup 213 during the spinning process.
[0110] The reagent mixer 9 furthermore comprises a housing 221
having sidewalls 222 surrounding at least the rim 214 of the mixing
cup 213 and thereby forming splash faces for collecting any liquid
flung out from the mixing cup 213. The housing also comprises a lid
223 for enclosing a space 224 surrounding the mixing cup 213 in
order to avoid reagent spills outside the space 224. The lid 223
has a central opening allowing reagents from the probe 10 to be
dispensed into the mixing cup 213 from above the reagent mixer 9 as
well as allowing the probe 10 to enter the mixing cup 213 for
collecting the mixed reagents.
[0111] According to a preferred embodiment the housing also
comprises a hose connection 227 for draining waste reagent or
cleansing liquid from the space 224, and a tap 226 is arranged for
dispensing cleansing liquid into the mixing cup 213 when
required.
[0112] In an alternative embodiment a releasable connection 212 is
provided between the probe 10 and the robot arm, enabling the
replacement of the probe 10 held by the robot arm by placing the
probe 10 in one of a number of free washing stations 8, where it is
released by the releasable connection 212, and where a new probe
10' is connected to the robot arm by means of the releasable
connection 212.
[0113] Having the appropriate input data, the control means of the
apparatus operates the robot arm to commence a staining or
treatment run by firstly moving the probe to a first reagent vial
3, into which the probe tip 211 is inserted into the liquid and is
aspirated into the probe 10 in an amount corresponding to the
number of samples to be stained or treated, in accordance with the
input data provided to the control means.
[0114] The probe 10 is subsequently, in a first operating mode
moved by the robot arm towards the slide rack system 5 in which the
slides 7 are mounted. The slides 7 are situated with the surface
horizontally oriented and the probe 10 releases the required amount
of reagent on the appropriate slides in accordance with the input
data. Alternatively, the probe 10 is in a second operating mode
moved by the robot arm towards the reagent mixer 9 where it
releases the reagent into the cup 213 of the reagent mixer 9, and
is subsequently moved to the probe washing station 8, where the
probe 10 is either washed or--in the alternative
embodiment--released into a free washing station 8, and another
probe situated in another washing station is connected to the robot
arm. The robot arm moves the new clean probe to a second selected
reagent vial 3 for collecting a selected amount of reagent from the
second vial 3, and the probe is thereafter by means of the robot
arm moved to the reagent mixer 9, where the reagent in the probe is
dispensed into the cup 13 of the mixer containing the first
selected reagent. The second operating mode can according to the
invention be commenced several times if more than two reagents are
to be mixed for a specific staining or treatment process.
[0115] The reagent mixer 9 mixes the reagents in the cup 213
thereof, and a clean probe 10 picked up from the washing station 8
by the robot arm is lowered into the cup 213 of the reagent mixer 9
to collect the mixed reagents, where after the robot arm moves the
probe 10 towards the second station 5 containing the slides 7, at
which the probe 10 releases the required amount of mixed reagent on
selected slides 7 in accordance with the input data.
[0116] The robot arm with probe 10 is subsequently directed to a
free washing station 8, and the probe 10 is either washed or
alternatively replaced by a clean probe, where after the process in
accordance with the first or the second operating mode may be
repeated or continued with a new reagent or reagent mixture.
[0117] The present invention is highly suitable for on-board mixing
in an automated cytological/IHC/ISH instrument.
[0118] It should be understood that by having an inner cup with
smooth surface and no sharp edged, cleaning and washing is made
easy. Other mixing devices using propellers or baffles or similar,
all have large surface areas, edges and concavities, whereto
reagents may adhere, whereby thorough washing becomes difficult,
possibly resulting in unwanted carry-over.
[0119] By design, the shape of the cup allows debris, fines,
precipitates or other solids of both high and low density to be
centrifuged up and out of the cup at high rotation speeds.
Advantageously the speed may be controlled by control means
operating according to the processing protocol carried out for the
samples present in the apparatus, i.e. if the reagents present in
the mixer cup have high viscosity it may be advantageous applying a
very high speed of rotation to ensure that all reagents are removed
from the cup.
[0120] Also, the shape of the cup allow for the specific separation
of immiscible liquid phases, by first removing the liquid with the
highest density during centrifugation. It should be understood that
the shape of the cup makes the mixing almost independent of the
volume. The mixing action is the same for a wide range of volumes
in the same cup. In contrast, the mixing action of e.g. a propeller
mixer is depending on e.g. the volume of reagents, propeller size
and container size.
[0121] The rotating action of the cup allows air to escape from the
mixture and prevent foaming. The rotating action can suppress the
formation and build-up of foam by forcing the air/liquid foam down
to the liquid surface.
[0122] Preferably the rotation pattern (speed and direction) is
controlled by controlling means, such as a computer, according to
the sample processing protocol, thereby optimising the quality the
mixing in the mixer cup in respect to the reagents present in the
mixer cup, e.g. in such manner that the formation and build up of
foam is suppressed.
[0123] The rotating action of the cup does not allow much air from
the atmosphere to be sucked down in the reagent mixture. This will
limit the pollution and e.g. oxidation of air sensitive reagents.
Other methods of mixing, like the action of propellers, can draw
air down into the liquid during mixture.
[0124] It should be understood that by changing the direction of
the rotation, the mixing action could be changed from being very
gentle to being very vigorous. Both changing the speed and
direction of the rotation and a combination of rotations with
variable speed, various speeds, stops and change of direction can
control the flow pattern in the cup.
[0125] Also, the cup can be covered with a neutral gas like the
noble gasses, nitrogen, and carbon dioxide or similar to further
protect the reagents during mixing. Preferable, the protective gas
is heavier than atmospheric gas. The layer of protective gas can
easily be added and removed, and can cover any of the volumes the
cup can hold.
[0126] By using an on board mixing device, the reagents mixing may
be done in a batch-mode to make mixtures for treating multiple
samples.
[0127] Also, the on board mixing allow for large dilution ratios
and the possibility to dilute or mix reagents, using only a small
portion of the mixture for treatment of samples, and discarding the
rest of the mixture. This is advantageous, as some dilutions ratios
are very large, and too small volumes of the concentrated reagent
cannot easily be accurately measured and added. Often a minimum
volume of reagent to be measured is defined. The on board cup
mixing design allow for a very high degree of flexibility in the
staining protocol.
[0128] As can be easily understood from the foregoing, the basic
concepts of the present invention may be embodied in a variety of
ways. It involves both mixing techniques as well as devices to
accomplish the appropriate mixer element. In this application, the
mixing techniques are disclosed as part of the results shown to be
achieved by the various devices described and as steps which are
inherent to utilization. They are simply the natural result of
utilizing the devices as intended and described. In addition, while
some devices are disclosed, it should be understood that these not
only accomplish certain methods but also can be varied in a number
of ways. Importantly, as to all of the foregoing, all of these
facets should be understood to be encompassed by this
disclosure.
[0129] The discussion included in this application is intended to
serve as a basic description.
[0130] The reader should be aware that the specific discussion may
not explicitly describe all embodiments possible; many alternatives
are implicit. It also may not fully explain the generic nature of
the invention and may not explicitly show how each feature or
element can actually be representative of a broader function or of
a great variety of alternative or equivalent elements. Again, these
are implicitly included in this disclosure. Where the invention is
described in device-oriented terminology, each element of the
device implicitly performs a function. Apparatus claims may not
only be included for the device described, but also method or
process claims may be included to address the functions the
invention and each element performs. Neither the description nor
the terminology is intended to limit the scope of the claims that
will be included in any subsequent patent application.
[0131] It should also be understood that a variety of changes may
be made without departing from the essence of the invention. Such
changes are also implicitly included in the description. They still
fall within the scope of this invention. A broad disclosure
encompassing both the explicit embodiment(s) shown, the great
variety of implicit alternative embodiments, and the broad methods
or processes and the like are encompassed by this disclosure and
may be relied upon when drafting the claims for any subsequent
patent application. It should be understood that such language
changes and broader or more detailed claiming may be accomplished
at a later date or in the event the applicant subsequently seeks a
patent filing based on this filing. With this understanding, the
reader should be aware that this disclosure is to be understood to
support any subsequently filed patent application that may seek
examination of as broad a base of claims as deemed within the
applicant's right and may be designed to yield a patent covering
numerous aspects of the invention both independently and as an
overall system.
[0132] Further, each of the various elements of the invention and
claims may also be achieved in a variety of manners. Additionally,
when used, the term "element" is to be understood as encompassing
individual as well as plural structures that may or may not be
physically connected. This disclosure should be understood to
encompass each such variation, be it a variation of an embodiment
of any apparatus embodiment, a method or process embodiment, or
even merely a variation of any element of these. Particularly, it
should be understood that as the disclosure relates to elements of
the invention, the words for each element may be expressed by
equivalent apparatus terms or method terms--even if only the
function or result is the same. Such equivalent, broader, or even
more generic terms should be considered to be encompassed in the
description of each element or action. Such terms can be
substituted where desired to make explicit the implicitly broad
coverage to which this invention is entitled. As but one example,
it should be understood that all actions may be expressed as a
means for taking that action or as an element which causes that
action. Similarly, each physical element disclosed should be
understood to encompass a disclosure of the action which that
physical element facilitates. Regarding this last aspect, as but
one example, the disclosure of a "mix element" should be understood
to encompass disclosure of the act of "mixing"--whether explicitly
discussed or not--and, conversely, were there effectively
disclosure of the act of "mixing", such a disclosure should be
understood to encompass disclosure of a "mix element" and even a
"means for mixing" Such changes and alternative terms are to be
understood to be explicitly included in the description.
[0133] Any patents, publications, or other references mentioned in
this application for patent are hereby incorporated by reference.
In addition, as to each term used it should be understood that
unless its utilization in this application is inconsistent with
such interpretation, common dictionary definitions should be
understood as incorporated for each term and all definitions,
alternative terms, and synonyms such as contained in the Random
House Webster's Unabridged Dictionary, second edition are hereby
incorporated by reference. Finally, all references listed in the
list of References To Be Incorporated By Reference or other
information statement filed with the application are hereby
appended and hereby incorporated by reference, however, as to each
of the above, to the extent that such information or statements
incorporated by reference might be considered inconsistent with the
patenting of this/these invention(s) such statements are expressly
not to be considered as made by the applicant(s).
[0134] Thus, the applicant(s) should be understood to have support
to claim and make a statement of invention: i) each of the mixing
devices as herein disclosed and described, ii) the related methods
disclosed and described, iii) similar, equivalent, and even
implicit variations of each of these devices and methods, iv) those
alternative designs which accomplish each of the functions shown as
are disclosed and described, v) those alternative designs and
methods which accomplish each of the functions shown as are
implicit to accomplish that which is disclosed and described, vi)
each feature, component, and step shown as separate and independent
inventions, vii) the applications enhanced by the various systems
or components disclosed, viii) the resulting products produced by
such systems or components, ix) each system, method, and element
shown or described as now applied to any specific field or devices
mentioned, x) methods and apparatuses substantially as described
hereinbefore and with reference to any of the accompanying
examples, xi) the various combinations and permutations of each of
the elements disclosed, and xii) each potentially dependent claim
or concept as a dependency on each and every one of the independent
claims or concepts presented.
[0135] With regard to claims whether now or later presented for
examination, it should be understood that for practical reasons and
so as to avoid great expansion of the examination burden, the
applicant may at any time present only initial claims or perhaps
only initial claims with only initial dependencies. Support should
be understood to exist to the degree required under new matter
laws--including but not limited to European Patent Convention
Article 123(2) and United States Patent Law 35 USC 132 or other
such laws--to permit the addition of any of the various
dependencies or other elements presented under one independent
claim or concept as dependencies or elements under any other
independent claim or concept. In drafting any claims at any time
whether in this application or in any subsequent application, it
should also be understood that the applicant has intended to
capture as full and broad a scope of coverage as legally available.
To the extent that insubstantial substitutes are made, to the
extent that the applicant did not in fact draft any claim so as to
literally encompass any particular embodiment, and to the extent
otherwise applicable, the applicant should not be understood to
have in any way intended to or actually relinquished such coverage
as the applicant simply may not have been able to anticipate all
eventualities; one skilled in the art, should not be reasonably
expected to have drafted a claim that would have literally
encompassed such alternative embodiments.
[0136] Further, if or when used, the use of the transitional phrase
"comprising" is used to maintain the "open-end" claims herein,
according to traditional claim interpretation. Thus, unless the
context requires otherwise, it should be understood that the term
"comprise" or variations such as "comprises" or "comprising", are
intended to imply the inclusion of a stated element or step or
group of elements or steps but not the exclusion of any other
element or step or group of elements or steps. Such terms should be
interpreted in their most expansive form so as to afford the
applicant the broadest coverage legally permissible.
[0137] Finally, any claims set forth at any time are hereby
incorporated by reference as part of this description of the
invention, and the applicant expressly reserves the right to use
all of or a portion of such incorporated content of such claims as
additional description to support any of or all of the claims or
any element or component thereof, and the applicant further
expressly reserves the right to move any portion of or all of the
incorporated content of such claims or any element or component
thereof from the description into the claims or vice-versa as
necessary to define the matter for which protection is sought by
this application or by any subsequent continuation, division, or
continuation-in-part application thereof, or to obtain any benefit
of, reduction in fees pursuant to, or to comply with the patent
laws, rules, or regulations of any country or treaty, and such
content incorporated by reference shall survive during the entire
pendency of this application including any subsequent continuation,
division, or continuation-in-part application thereof or any
reissue or extension thereon.
* * * * *