U.S. patent application number 10/486932 was filed with the patent office on 2006-09-21 for fixtures for use in parallel processing bio-chips.
Invention is credited to CliffordL Anderson, RichardD JR. Belval, RichardE Maldonado.
Application Number | 20060210451 10/486932 |
Document ID | / |
Family ID | 26910606 |
Filed Date | 2006-09-21 |
United States Patent
Application |
20060210451 |
Kind Code |
A1 |
Anderson; CliffordL ; et
al. |
September 21, 2006 |
Fixtures for use in parallel processing bio-chips
Abstract
A holder (10) housing defines a plurality of individual wells
(14) sized to retain a substrate such as a bio-chip (20) or
microscope slide in a vertical orientation, and to retain a volume
of liquid (22) sufficient to immerse the bio-chip. The wells are
spaced-apart with an approximately 9 mm pitch and the holder has a
form factor approximating an SBS Standard 96 well microplate. The
holder configuration is such that it can be manipulated with
standard robotic equipment (30), and can be fabricated using
injection molding processes.
Inventors: |
Anderson; CliffordL; (Tempe,
AZ) ; Maldonado; RichardE; (Fountain Hills, AZ)
; Belval; RichardD JR.; (New Fairfield, CT) |
Correspondence
Address: |
GE HEALTHCARE BIO-SCIENCES CORP.;PATENT DEPARTMENT
800 CENTENNIAL AVENUE
PISCATAWAY
NJ
08855
US
|
Family ID: |
26910606 |
Appl. No.: |
10/486932 |
Filed: |
August 16, 2002 |
PCT Filed: |
August 16, 2002 |
PCT NO: |
PCT/US02/26078 |
371 Date: |
January 6, 2005 |
Current U.S.
Class: |
422/400 |
Current CPC
Class: |
Y10T 436/11 20150115;
B01L 9/52 20130101; G01N 2035/00089 20130101; B01L 2300/04
20130101; B01L 2300/0819 20130101; G01N 1/312 20130101; B01L 3/5085
20130101; G01N 2035/00138 20130101; G02B 21/26 20130101; Y10T
436/112499 20150115; B01L 2300/0822 20130101; G01N 2035/00158
20130101 |
Class at
Publication: |
422/104 |
International
Class: |
B01L 9/00 20060101
B01L009/00 |
Foreign Application Data
Date |
Code |
Application Number |
Aug 16, 2001 |
US |
60312719 |
Aug 9, 2002 |
US |
10216057 |
Claims
1. A holder to receive a plurality of substrates selected from a
group consisting of bio-chips and microscope slides, the holder
comprising: a holder housing whose upper region defines a plurality
of separate channels, each of said channels sized to receive one of
said bio-chips in a vertical orientation and to retain an amount of
liquid sufficient to immerse a received bio-chip; said holder
having a form factor sized to accommodate manipulation by a robotic
mechanism.
2. The holder of claim 1, wherein an upper surface of holder
conforms to an SBS Standard 96-well microplate.
3. The holder of claim 1, wherein said channels are defined with a
spaced-apart pitch approximating 9 mm.
4. The holder of claim 1, wherein each of said substrates is a
bio-chip having a form factor approximating a standard microscope
slide.
5. The holder of claim 1, further including a lid member, sized to
sealingly attach to an upper surface of said holder.
6. The holder of claim 1, wherein said amount of liquid is less
than about 4 ml.
7. The holder of claim 1, wherein said amount of liquid is about
3.4 ml.
8. The holder of claim 1, wherein said holder is injection
molded.
9. A robotically-manipulatable kit for use with substrates selected
from a group consisting of bio-chips and microscope slides,
comprising at least two of: a holder having a form factor sized to
accommodate manipulation by a robotic mechanism and including a
housing whose upper region defines a plurality of separate channels
spaced-apart from each other with a pitch P, each of said channels
sized to receive one of said substrates in a vertical orientation
and to retain an amount of liquid sufficient to immerse a received
substrate; a reservoir having a form factor sized to accommodate
manipulation by a robotic mechanism and sized to receive said
holder; and a positioning tool having a form factor sized to
accommodate manipulation by a robotic mechanism and sized to retain
a plurality of said substrates with said pitch P.
10. The kit of claim 9, wherein an upper surface of holder conforms
to an SBS Standard 96-well microplate.
11. The kit of claim 9, wherein said pitch P is about 9 mm.
12. The kit of claim 9, wherein each of said substrates is a
bio-chip having a form factor approximating a standard microscope
slide.
13. The kit of claim 9, further including a lid member, sized to
sealingly attach to an upper surface of said holder.
14. The kit of claim 9, wherein volume of liquid retainable in each
said channel is less than about 4 ml.
15. The kit of claim 9, wherein volume of liquid retainable in each
said channel is about 3.4 ml.
16. The kit of claim 9, wherein said holder is injection
molded.
17. A robotically-manipulatable kit for use with microscope-sized
slides, comprising at least two of: a holder having a form factor
sized to accommodate manipulation by a robotic mechanism and
including a housing whose upper region defines a plurality of
separate channels spaced-apart from each other with a pitch P, each
of said channels sized to receive one of said slides in a vertical
orientation and to retain an amount of liquid sufficient to immerse
a received slide; a reservoir having a form factor sized to
accommodate manipulation by a robotic mechanism and sized to
receive said holder; and a positioning tool having a form factor
sized to accommodate manipulation by a robotic mechanism and sized
to retain a plurality of said slides with said pitch P.
18. The kit of claim 17, wherein said slides include at least one
slide selected from a group consisting of (a) a slide of cytology
staining, (b) a slide for parasite staining, (c) and a slide for
culturing cells.
19. The kit of claim 17, wherein an upper surface of holder
conforms to an SBS Standard 96-well microplate.
20. The kit of claim 17, wherein said pitch P is about 9 mm.
21. The kit of claim 17, further including a lid member, sized to
sealingly attach to an upper surface of said holder.
22. The kit of claim 17, wherein volume of liquid retainable in
each said channel is less than about 4 ml.
23. The kit of claim 17, wherein volume of liquid retainable in
each said channel is about 3.4 ml.
24. The kit of claim 17, wherein said holder is injection molded.
Description
PRIORITY CLAIM
[0001] Priority is claimed to co-pending U.S. provisional patent
application Ser. No. 60/312,719, filed on 16 Aug. 2001 entitled
"Fixtures for Use in Parallel Processing Bio-Chips".
FIELD OF THE INVENTION
[0002] The invention relates generally to fixtures that hold
bio-chips, microscope slides and the like, and more particularly to
fixtures that can be manipulated either manually or robotically
during parallel processing of bio-chips, including bio-chips in
microscope-slide format.
BACKGROUND OF THE INVENTION
[0003] A variety of bio-chips is known in the art. Typically a
bio-chip includes a substrate upon which an array of test sites may
be defined, the number of sites ranging from a thousand to ten
thousand or more on a bio-chip that measures perhaps a few cm by a
few cm. Various experiments may be carried out at the various
sites, e.g., using reagents, and frequently the bio-chip will be
moved, manually or robotically during experiments or testing.
[0004] Robotic repositioning of bio-chips is preferred in that such
manipulation is generally more accurate than manual repositioning,
and can be carried out more rapidly. Understandably more rapid
manipulation of bio-chips is desired in that more experiments can
be carried out per unit time.
[0005] Although several prior art holders can retain multiple
bio-chips, such holders frequently have shortcomings. For example,
Marsh Bio Products, Inc. of Rochester, N.Y. 14610 produces the
so-called DR1205 reservoir. This holder provides channels for
twelve slides but requires a relatively high volume, 5 ml, of
reagent per channel. Since cost of reagents can be high in various
experiments, it is desired to reduce the amount of reagent needed
per retained bio-chip. Other manufacturers provide holders to
retain bio-chips, which holders are fabricated from expensive
material, for example stainless steel. Too often, prior art
bio-chip holders fail to fully protect the active surface of the
bio-chip. Understandably, failing to adequately protect the
bio-chip can affect the nature and quality of the test results.
Various prior art bio-chip holders are configured such that robotic
manipulations including microtiter formatting, automatic
filling/draining of channels, automatic stacking of holders cannot
readily be accomplished. Further, various prior art bio-chip
holders do not provide an independent channel for each retained
bio-chip, which omission can result in cross-contamination.
[0006] As such, there is a need for a fixture that can retain one
or more bio-chips during various phases of experimentation, which
fixture permits robotic movement such that parallel processing of
bio-chips retained therein can be carried out. Further, such
fixture preferably should provide an independent channel or well
for retained bio-chips to minimize cross-contamination, and
preferably should require a relatively small amount of reagent per
bio-chip. Such holder should provide a mechanism by which each well
may rapidly be filled or emptied with a liquid of interest. Such
fixture preferably should be made of a relatively inexpensive
material, and should robustly protect bio-chips retained
therein.
[0007] The present invention provides such a fixture, and a method
of retaining bio-chips while permitting manual and/or robotic
manipulation of the bio-chips. Further the present invention can
accommodate standard glass microscope slides as well and can
facilitate manual and/or robotic manipulation of such slides for a
variety of applications.
SUMMARY OF THE INVENTION
[0008] A holder according to the present invention includes a
housing that defines a plurality of individual wells or channels
for retaining a liquid and a bio-chip intended to react in some
manner with the liquid. The holder and each well is sized to retain
a relatively minimal amount of liquid, while ensuring that a
bio-chip inserted into the well is fully immersed. Inter-well
fluid-communication does not occur and resultant
cross-contamination is thus reduced. The holder configuration is
preferably standardized in size and form factor such that
conventional microscope slide-sized bio-chips are retained, and the
holder itself may be manipulated with standard robotic mechanisms.
Further, the holder is sized to be sealingly covered with standard
lids, or with tools according to the present invention. The various
holders and tools preferably are configured to be fabricated from
durable material, using standardized techniques such as injection
molding. Tools and reservoirs for use with an array of bio-chips
are also described. The holders may also receive standard
microscope slides, enabling the present invention to be used in
applications including slide staining, e.g., for histology,
cytology, etc.
[0009] Other features and advantages of the invention will appear
from the following description in which the preferred embodiments
have been set forth in detail, in conjunction with the accompanying
drawings.
BRIEF DESCRIPTION OF THE DRAWINGS
[0010] FIG. 1 is a perspective plan view of a first embodiment of a
holder or bio-array rack for bio-chips, according to the present
invention. This is generally described herein as a holder that is
part of a two component system;
[0011] FIG. 2 is a perspective plan view of a preferred embodiment
of a bio-array tool and a holder, loadable with bio-chips using the
bio-array tool, according to the present invention;
[0012] FIG. 3 is a perspective plan view of an embodiment of a
reagent reservoir holder for receiving bio-chips, according to the
present invention. This is part of a two component system;
[0013] FIG. 4 is a perspective plan view of an alternative
embodiment of a reagent reservoir holder for receiving bio-chips,
according to the present invention;
[0014] FIG. 5 is a perspective plan view of a fifth embodiment of a
holder for bio-chips, according to the present invention;
[0015] FIG. 6 is a perspective plan view of a tool for removing an
array of bio-chips from a holder, according to the present
invention; and
[0016] FIG. 7 is a perspective plan view showing a bio-chip holder
and reagent reservoir, according to the present invention, and a
robotic mechanism.
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT
[0017] The present invention provides a variety of holders and
tools for the manipulation of a plurality of bio-chips for
hybridization, washing and analysis steps. In particular, the
holders and tools of the invention find use in assays utilizing
bio-chips such as those described in PCT US 00/34145 and PCT
US01/02664, both of which are expressly incorporated by reference
in their entirety. These bio-chips comprise a rigid support with a
flexible membrane top that encompasses the reaction chamber
comprising the array of capture biomolecules, particularly nucleic
acids. The chips comprise ports for the introduction of samples
comprising the analytes of interest. After introduction of the
sample, the bio-chips are incubated for a period of time sufficient
to allow binding (referred to as hybridization when the target
analyte and capture probes are nucleic acids), frequently with
mixing. Washing solutions can be added with further incubation,
although as described herein, the washing steps may occur later.
The flexible membrane is generally removed, and then further
optional washing steps can be done, followed by detection of the
target analytes, usually by insertion of the bio-chips into a
scanner or reader.
[0018] Accordingly, there is a need for holders and tools to allow
the simultaneous manipulation of a plurality of bio-chips to reduce
time, cost, labor and increase reproducibility and accuracy.
[0019] Thus, the present invention provides a number of holders and
tools to facilitate this manipulation. As described below, in one
embodiment, the holder comprises slots into which the bio-chips are
inserted, with an optional top. This holder is then inserted into a
reservoir system (e.g. a two component system), that can include
either a single large reservoir, such that the bio-chips are in
fluid communication, or separate reservoirs, such that the
bio-chips are fluidically isolated. Alternatively, the holder can
comprise both slots for the insertion of the chips, as well as the
reservoir (e.g. a one component system). In addition, tools are
provided that allow ease of handling; for example, in a two
component system, a removal tool (as depicted in FIG. 6) can be
used to "pop" out the chips for transfer to other systems such as a
detection system.
[0020] FIG. 1 depicts a first embodiment of a bio-chip holder or
bio-array rack 10 comprising a housing surface 12 in which a
plurality of bio-chip retaining channels or reservoirs 14 are
formed. Preferably first and second sidewalls 16 are affixed to
surface 12, and include indicia 18 uniquely identifying each
channel. For example in FIG. 1, holder 12 provides twelve channels,
for which indicia 18 includes numerals 1, 2, . . . 12. The use of
indicia 18 can reduce human error when dealing with bio-chips 20
inserted into or removed from a given channel. In FIG. 1, bio-chip
20-12 is shown somewhat removed from underlying channel number 12.
By contrast, channel number 9 is shown with a bio-chip 20-9
substantially within the channel. One or more reagents 22 may be
present in each channel 14-x, such that sites on a bio-chip 20-x
within a channel will be exposed to such reagents. Preferably
channels defined in holder 10 permit completely immersing a
bio-chip with reagent 22 within the channel. In the preferred
embodiments described herein, each channel 14-x is sized to receive
a standard 1'' (2.54 cm).times.3'' (7.62 cm).times.0.04'' (1.3 mm)
microscope slide, which is to say, each bio-chip 20-x preferably is
the size of a standard microscope slide. Thus, components 20-x
could, if desired, be microscope slides. It is understood that
holder 10 could of course include channels of varying size and
volume and configuration. In a preferred embodiment, the volume of
liquid used within each well for an associated bio-chip can be as
little as about 3.4 ml. As noted since many reagents are costly,
the ability to perform tests with a reduced volume of liquid can be
very advantageous.
[0021] Preferably the outer dimensions and form factor of holder
10, and indeed each holder or tool or reservoir embodiment
described herein, is such that the holders conform substantially
closely to SBS Standard 96-well microplate dimensions. Such
standardization of the form factor of holders according to the
present invention permits existing robotic-type mechanisms,
depicted generically by arm 30, to manipulate the holders and
bio-chips (or microscope slides) within. (See also FIG. 7.)
Further, it is preferred that the spaced-apart pitch P between
adjacent channels or wells 14-x in holder 10 is a standard
dimension, for example a 9 mm pitch corresponding to multiple-pipet
dispensing port pitches. The ability of each holder 10 to be
accurately and rapidly manipulated by mechanism 30 permits holding
and moving a plurality of retained bio-chips, for example from one
reagent reservoir to another reservoir. In the configuration shown
in FIG. 1, gravity advantageously assists in the process of loading
each bio-chip 20-x into a corresponding well or channel 14-x in
holder 10. Further, vertical grooves 32 defined in the vertical
edges of each channel help retain an associated bio-chip such that
the vertical surfaces (in FIG. 1) of the bio-chip do not contact
the vertical walls of the well within housing 10. Further, the well
configuration preferably is symmetrical such that a bio-chip can be
inserted without regard to orientation, while ensuring full
immersion of the relevant bio-chip surfaces in reagent or other
liquid 22.
[0022] Automation using the present invention is further promoted
by providing holder 10 with at least one input port 32 and at least
one output port 34. Input port 32 is preferably in one-way fluid
communication with each channel or well 14-x in the holder, and
facilitates rapidly filling each well with a liquid. Each well
advantageously has a level marker 36 such that the correct fill
level can be rapidly and visually confirmed. Using simple valves or
the like, fluid communication is one-way such that liquid contents
of one well do not communicate with the liquid contents of another
well. Output port 34 can be used to rapidly purge liquid from each
of the wells. Further, the form factor of each holder according to
the present invention permits the holders to be stacked, and to be
sealingly covered using a standard microplate lid.
[0023] Holder 10 may be fabricated in a variety of ways using any
of several materials. However in a preferred embodiment, holder 10
is injection moldable to reduce manufacturing costs. Holder 10
preferably is fabricated from a material that is resistant to
chemicals, for example, an acetyl copolymer material. Such material
advantageously can tolerate a wide temperature range and can be
easily cleaned, and in fact is dishwasher safe. Further, such
material (or other material) from which holder 10 is fabricated
preferably will be substantially opaque to optical energy
encountered during light-sensitive testing or experimental phases
of the bio-chips retained within.
[0024] FIG. 2 depicts a preferred embodiment of a bio-array
positioning tool 40 that is sized to retain a plurality of
bio-chips 20-x with pitch P to permit tool 40 to be lowered over
the upper surface of a holder, e.g. holder 10 such that each
bio-chip is lowered into an associated well or channel 14 in holder
10 and is fully seated therein. (Unless otherwise noted, like
reference numerals in one figure herein may be understood to refer
to similar or identical elements.) The lower surface 42 of tool 40
preferably includes a mechanism 44 for retaining each bio-chip 20-x
within the tool. Without limitation, mechanism 44 may include a
groove formed in lower surface 44 to fictionally retain an edge of
each bio-chip to be held by tool 40. Use of tool 40 can
advantageously promote proper seating of bio-chips 20-x in holder
10 such that full immersion with reagent or other liquid 22
results. Further, tool 40 is convenient for at least partially
removing the array of bio-chips from holder 10. Tool 40 preferably
is fabricated using techniques and materials similar to what has
been described above with respect to holder 10. As tool 40 and
holder 10 preferably are uniformly sized, tool 40 can be used as a
sealing lid to cover the upper surface of holder 10, with reagent
and bio-chips within the holder. In a closed disposition, tool 40
preferably matingly seals to the holder 10 such that a robotic
mechanism, e.g., mechanism 30 in FIG. 1, can manipulate holder 10,
tool 40, and the bio-chips within.
[0025] FIG. 3 depicts what may be termed a reservoir holder 50 that
defines a plurality of individual channels or wells 14-1, 14-2, . .
. 14-N, each sized to retain a liquid or reagent 22 and to receive
a bio-chip such as bio-chip 20-x, described above with reference to
FIGS. 2 and 3. Holder 50 includes four sides 52, a bottom 54, and a
top member 56 that defines individual wells or channels 14-x.
Again, it is understood that holder 50 is sized such that it may be
sealingly covered with a standard lid including tool 40. If
desired, each channel or well 14-x formed in upper member 56 may
include an enlarged end region 58 to promote more rapid filling
with liquid or reagent 22. It is understood that in the preferred
embodiment there is no cross-contamination from fluid in one
channel with fluid in another channel in holder 50.
[0026] FIG. 4 depicts a reservoir holder 60 similar to holder 50
shown in FIG. 3, except that optionally a larger volume of reagent
22 may be retained within. FIG. 5 depicts an embodiment of a
reservoir holder 70 that preferably retains more liquid or reagent
22 than does holder 60 or holder 50. Reservoir holder 70 comprises
four side walls 72, and bottom member 74 and defines an open volume
76 within that may be filled with liquid or reagent 22. Once filled
with a liquid, tool 40 (or the like) loaded with bio-chips 20-x may
be lowered over holder 70 such that each bio-chip is immersed in
the liquid retained within volume 76. Holder 70 preferably includes
at least one input port 78 and at least one output port 80 to
facilitate the rapid filling and draining of volume 76 with liquid
22. Port 80 may be used, for example, as a bleed opening in a water
bath phase of parallel-processing of bio-chips. Again, it is
understood that holder 70 like the other holders described herein
is sized such that a standard lid including tool 40 may be sealing
attached to the top portion of the holder. Further, the
standardized form factor of the various holders described herein is
such that holder 70 may be manipulated with a robotic mechanism
such as mechanism 30 (see FIG. 1).
[0027] Turning now to FIG. 6, a bio-array removal tool 90 is
depicted. Removal tool 90 includes a base member 92, two
spaced-apart short sidewall members 9 and two spaced-apart longer
sidewall members 96. An upper ledge formed on each of the long
sidewall members 96 defines a series of notches 98-x, preferably
spaced-apart with the same pitch P as was defined with regard to
FIGS. 1-4. Notches 98-x in removal tool 90 can frictional engage
respective edges of bio-chips 20-x for purposes of at least
partially withdrawing the bio-chips from a holder, for example
holder 10 in FIG. 1. It is understood that preferably removal tool
90 may be fabricated using techniques and materials similar to that
used for the various embodiments of tools and holders described
above herein.
[0028] FIG. 7 shows a robotic mechanism 7 in juxtaposition over a
reservoir such as reservoir 50 described with respect to FIG. 3,
and an overlying holder such as holder 10 described with respect to
FIG. 1. Preferably the spaced-apart distance D between arms of the
robotic mechanism is such that the mechanism can fit within the
preferably about 1.5 mm or about 3 mm gap clearance provided on
either side of holder 10 and reservoir 50. This gap clearance helps
minimize capillary action that could cause fluid to cross between
channels, such as channels 14-x, described earlier herein. The
modular-like design of the present invention is such that when
positioning tool 40 (see FIG. 2) is inserted into a holder, e.g.,
10, robotic mechanism 30 can be controlled to selectively
manipulate, e.g., pick-up, only positioning tool 40, or positioning
tool 40 and holder 10, or to pick-up positioning tool 40, holder
10, and reservoir 50.
[0029] In summary, the various embodiments of the present invention
provide holders or fixtures, reservoirs, and tools that can
expedite the parallel processing of bio-chips. In contrast to prior
art mechanisms, the described embodiments lend themselves to
robotic manipulation and are useable with standardized components
such as lids, and microscope slide-sized bio-chips. While exemplary
dimensions and form factors have been described, it is understood
that other dimensions and shapes could instead be used.
[0030] In addition, as will also be appreciated by those in the
art, the holders, tools and biochips of the invention may be part
of high throughput screening (HTS) system utilizing any number of
components. Fully robotic or microfluidic systems include automated
liquid-, particle-, cell- and organism-handling including high
throughput pipetting to perform all steps of gene targeting and
recombination applications. This includes liquid, particle, cell,
and organism manipulations such as aspiration, dispensing, mixing,
diluting, washing, accurate volumetric transfers; retrieving, and
discarding of pipet tips; and repetitive pipetting of identical
volumes for multiple deliveries from a single sample aspiration.
These manipulations are cross-contamination-free liquid, particle,
cell, and organism transfers. This instrument performs automated
replication of microplate samples to filters, membranes, and/or
daughter plates, high-density transfers, full-plate serial
dilutions, and high capacity operation.
[0031] The system used can include a computer workstation
comprising a microprocessor programmed to manipulate a device
selected from the group consisting of a thermocycler, a
multichannel pipettor, a sample handler, a plate handler, a gel
loading system, an automated transformation system, a gene
sequencer, a colony picker, a bead picker, a cell sorter, an
incubator, a light microscope, a fluorescence microscope, a
spectrofluorimeter, a spectrophotometer, a luminometer, a CCD
camera and combinations thereof.
[0032] It is also to be understood that the present invention may
be used in various manipulation of microscope slide-sized slides
aside from bio-chips. Without limitation, exemplary other such uses
could include ctyology staining, parasite staining, and various
cell culture techniques.
[0033] Modifications and variations may be made to the disclosed
embodiments without departing from the subject and spirit of the
invention as defined by the following claims.
* * * * *