United States Patent [19J
`Zanzucchi et al.
`
`I lllll llllllll Ill lllll lllll lllll lllll lllll lllll lllll lllll llllll Ill lllll llll
`US005585069A
`5,585,069
`[11] Patent Number:
`[ 45] Date of Patent:
`Dec. 17, 1996
`
`[54] PARTITIONED MICROELECTRONIC AND
`FLUIDIC DEVICE ARRAY FOR CLINICAL
`DIAGNOSTICS AND CHEMICAL SYNTHESIS
`
`[75]
`
`Inventors: Peter J. Zanzucchi, Lawrenceville;
`Satyam C. Cherukuri, Cranbury;
`Sterling E. McBride, Lawrenceville, all
`of N.J.
`
`[73] Assignee: David Sarnoff Research Center, Inc.,
`Princeton, N.J.
`
`[21] Appl. No.: 338,703
`
`[22]
`
`Filed:
`
`Nov. 10, 1994
`
`[51]
`[52]
`
`Int. Cl.6
`.... . ....... . .................. BOIL 3/00; GOIN 35/00
`U.S. Cl . ........................... 422/100; 422/58; 422/68.1;
`436/43; 204/450; 204/600
`[58] Field of Search ................................ 204/403, 299 R,
`204/180.1, 454, 452, 601, 603; 436/43;
`935/87, 88; 422/68.1
`
`[56]
`
`References Cited
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`
`Primary Examiner-Kathryn Gorgos
`Assistant Examiner-Alex Noguerola
`Attorney, Agent, or Firm-William J. Burke
`
`[57]
`
`ABSTRACT
`
`A system for processing a plurality of tests or syntheses in
`parallel comprising a sample channel for moving samples
`into a microlaboratory array of a plurality of wells connected
`by one or more channels for the testing or synthesis of
`samples, a station for housing the array and an optical
`system comprising at least one light source and at least one
`light detector for measuring the samples in the array, and a
`means of electrically connecting the array to an apparatus
`capable of monitoring and controlling the flow of fluids into
`the array.
`
`Samples are loaded from a common loading channel into the
`array, processed in the wells and measurements taken by the
`optical system. The array can process many samples, or
`synthesize many compounds in parallel, reducing the time
`required for such processes.
`
`23 Claims, 10 Drawing Sheets
`
`THERMO FISHER EX. 1044
`
`
`

`

`5,585,069
`Page 2
`
`U.S. PATENT DOCUMENTS
`
`4,3 85,115
`4,683,914
`4,753,775
`4,756,884
`4,891,120
`4,908, 112
`4,948,961
`4,960,486
`4,963,498
`4,999,283
`4,999,284
`4,999,286
`5,000,817
`5,001,048
`5,006,749
`5,063,081
`5,066,938
`5,073,029
`5,118,384
`5,129,262
`5,140,1 61
`5,141,868
`5,143,854
`5,1 44,139
`5,147,607
`5,156,810
`5,164,598
`5,176,203
`5,178,190
`5,188,963
`5,189,914
`5,194,133
`5,200,051
`5,204,525
`5,212,988
`5,220,189
`5,229,297
`5,230,864
`5,238,853
`5,241,363
`5,250,263
`5,252,743
`5,262,127
`5,288,463
`5,296,114
`5,296,375
`5,304,487
`5,312,590
`5,322,258
`5,324,483
`5,324,633
`5,359,115
`5,384,261
`5,412,087
`5,420,328
`5,424,186
`5,427,946
`5,463,564
`5,480,614
`
`511983 Zabala et al. ............................. 435/33
`811987 Brisland ............................. 137/625.48
`6/1988 Ebersole et al ........................... 422/81
`711988 Hillman et al ............................ 422173
`111990 Sethi et al . .......................... 204/299 R
`3/1990 Pace .. ..................... ........ ..... 204/299 R
`8/1990 Hillman et al ....................... 250/252.l
`1011990 Perkins et al. ..... ..... ................ 156/633
`1011990 Hillman et al ............................ 436/69
`3/1991 Zavos et al ................................. 435/2
`311991 Ward et al. ................................. 435/4
`3/1991 Gawel et al .
`.......................... 43517.32
`3/1991 Aine ........................................ 156/633
`............................... 435/4
`3/1991 Taylor et al .
`411991 White et al. ............................ 310/323
`1111991 Cozzette et al . ............................ 427/2
`1111991 Kobashi et al ..................... 338/22 SD
`1211991 Eberly et al. ........................... 356/432
`6/1992 Harmon et al .......................... 156/643
`711992 White et al .
`.............................. 73/599
`811992 Hillman et al. ............ ............. 250/341
`8/1992 Shanks et al ........................... 435/288
`9/1992 Pirrung et al ........................... 436/518
`9/1992 Hillman et al. ......................... 250/341
`9/1992 Mochida ................................... 422/57
`10/1992 Ribi ...................................... 422/82.01
`11/1992 Hillman et al .......................... 250/341
`1/1993 Larzul ....................................... 165/61
`1/1993 Mettner .............................. 137/625.65
`211993 Stapleton ................................ 435/299
`3/1993 White et al. .............................. 73/599
`3/1993 Clark et al .......................... 204/299 R
`4/1993 Cozzette et al ......................... 204/403
`411993 Hillman et al ....................... 250/252.1
`5/1993 White et al. .............................. 73/599
`611993 Higashi et al .......................... 257/467
`711993 Schnipelsky et al .
`.................... 436/94
`711993 Columbus ............................... 422/100
`811993 Calzi et al. ............................... 436/68
`8/1993 Gamer .................................... 356/326
`10/1993 Manz ........................................ 422/81
`10/1993 Barrett et al ......................... 548/303.7
`1111993 Wise et al ................................. 422/98
`2/1994 Chemelli ................................... 422/58
`3/1994 Manz ................................... 204/180.1
`3/1994 Kricka et al . .... ...................... . 435/291
`4/1994 Wilding et al .......................... 435/291
`511994 Gunasingham ... ........................ 422/56
`611994 Bosch et al. .............................. 251/65
`6/1994 Cody et al. . ............................ 422/131
`6/1994 Foder et al .
`................................ 435/6
`10/1994 Campbell et al .
`...................... 558/110
`111995 Winkler et al .......................... 436/518
`511995 McGall et al . ..................... .... 536/24.3
`5/1995 Campbell ................................ 538/110
`6/1995 Fodor et al. ............................... . 435/6
`6/1995 Kricka et al . ........................... 435/291
`10/1995 Agrafiotis et al . ...................... 364/496
`111996 Kamahori .................................. 422/98
`
`OTHER PUBLICATIONS
`
`Fisher, "Microchips for Drug Compounds", New York
`Times, Mar. 3, 1991.
`The Economist, "The silver shotguns'', Dec. 14, 1991.
`Harrison et al, "Capillary Electrophoresis .. . ", Anal. Chem.
`vol. 64, 1992, pp. 1926--1932 Month not available.
`Mehregany,
`"Microelectromechanical Systems", 1993
`IEEE, Circuits & Devices, pp. 14-22 Month not available.
`
`Harrison et al, "Micromachining a Miniaturized . . . "
`Science, vol. 261 , Aug. 13, 1993, pp. 895-897.
`Fan et al, "Micromachining of Capillary . . . "Anal. Chem.
`vol. 66, No. 1, Jan. 1, 1994, pp. 177-184.
`Jacobson et al, "High Speed Separations ... ", Anal. Chem.
`vol. 66, 1994, pp. 1114-1118 month not available.
`Jacobson et al, "Effects of Injection . . . ",Anal. Chem. vol.
`66, No. 7, Apr. 1, 1994 month not available.
`Jerman et al, "Understanding Microval ve Technology", Sen­
`sors, Sep. 1994, pp. 26-36.
`Dasgupta et al, "Electroosmosis: A Reliable Fluid .. . ",
`Anal. Chem. vol. 66, No. 11 , Jun. 1, 1994, pp. 1792-1798.
`Wooley et al, "Ultra-high speed DNA . . . ", Proc. Natl.
`Acad. Sci. USA, vol. 91 , No. 1994, pp. 11348-11352 month
`unavailable.
`Dialog Search, May 19, 1994.
`Gazette entry for Semancik, et al., Temperature-Controlled,
`Micro machined Arrays for Chemical Sensor Fabrication and
`Operation, U.S. Patent No. 5,345,213 , Sep. 6, 1994.
`Gazette entry for Murphy, et al., Automated Capillary Scan­
`ning System, U.S. Patent No. 5,009,503, Apr. 23, 1991.
`Search Report for WO 93/22421 , Micro-Fabricated Sperm
`Handling Device, Nov. 11 , 1993.
`Search Report WO 93/22055, Fluid Handling in Micro-Fab­
`ricated Analytical Devices, Nov. 11, 1993.
`Search Report for WO 93/22054, Analysis Based on Flow
`Restriction, Nov. 11, 1993.
`Search Report for WO 93/22053, Micro-Fabricated Detec­
`tion Structures, Nov. 11 , 1993.
`Harmon, et al., Selectively in Electrophoretically Mediated
`Microanalysis by Control of Product Detection Time, Anal.
`Chem. 66:3797-3805, 1994.
`Patterson, et al., Electrophoretically Mediated Microanaly­
`sis of Calcium, J. Chromatog. A, 662:289-395, 1994.
`Excerpt for Chemical & Engineering News, Microfabricated
`Device is Chemistry Lab on a Chip, Dec. 12, 1991.
`Jacobson, et al., Precoloun Reactions with Electro Phoretic
`Analysis
`Integrated on a Microchip, Anal. Chem.
`66:4127-4132, 1994.
`Harmon, et al., Mathematical Treatment of Electrophoreti­
`cally Mediated Microanalysis, Anal. Chem. 65:2655-2662,
`1993.
`Avila Whitesides, Catalytic Activity of Native Enzymes
`During Capillary Electrophoresis: An Enzymatic Microre­
`actor, J. Org. Chem. 58:5508-5512, 1993.
`Harmon, et al., Electrophoretically Mediated Microanalysis
`of Ethanol, J. Chrom. A, 657:429-434, 1993.
`Bao and Regnier, Ultrarnicro Enzyme Assays in a Capillary
`Electrophoretic System, J. Chrom. 608:217-224, 1992.
`Richeter, et al., A Micromachined Electrohydrodynarnic
`(EHD) Pump, Sensors and Actuators A, 29:159-168, 1992.
`Bart, et al., Microfabricated Electrohydrodynarnic Pumps,
`Sensors and Actuators, A21-A23:193-197, 1990.
`Melcher, Traveling-Wave Induced Electroconvection, The
`Physics of Fluids, 9:1548-1555, 1966.
`Pickard, Ion Drag Pumping. I. Theory, J. Applied Physics
`34:246--250, 1963.
`Pickard, Ion Drag Pumping. II. Experiment, J. Applied
`Physics, 34:251-258, 1963.
`Stuetzer, Ion Drag Pumps, J. Applied Physics, 31:136--146,
`1960.
`Tracey, et al., Microfabricated Microhaemorheometer, pp.
`82-84, 1991.
`Medynski, Synthetic Peptide Combinatorial Libraries, Bio/
`Technology, vol. 12, Jul. 1994.
`
`THERMO FISHER EX. 1044
`
`
`

`

`5,585,069
`Page 3
`
`Jacobson, et al., Effects of Injection Schemes and Column
`Geometry on the Performance of Microchip Electrophoresis
`Devices, Anal. Chem. 1994, 66:1107-1113.
`Jacobson, et al., High-Speed Separations on a Microchip,
`anal. Chem. 1994, 66:1114-1118.
`Fan, et al., Micromachining of Capillary Electrophoresis
`Injectors and Separators on Glass Chips and Evaluation of
`Flow at Capillary Intersections, Anal. Chem., 1994,
`6:177-184.
`Megregany, Microelectromechanical Systems, Circuits and
`Devices, Jul. 1993.
`Harrison, et al., Micromachining a Miniaturized Capillary
`Electrophoresis-Based Chemical Analysis System on a
`Chip, Science, vol. 261, Aug. 13, 1993.
`Harrison, et al., Capillary Electrophoresis and Sample Injec­
`tion Systems Integrated on a Planar Glass Chip, Anal. Chem.
`1992, 64: 1926-1932.
`
`Fisher, Microchips for Drug Compounds, New York times,
`Mar. 3, 1991.
`Fodor, ete al., Light-Directed, Spatially Addressable Paral­
`lel Chemical Synthesis, Research Article, Science, vol. 251,
`Feb. 15, 1991, pp. 767-773.
`The Silver Shotguns, The Economist, Dec. 14-20, 1991.
`Howe, et al., Silicon Micromechanics; Sensors and Actua­
`tors on a Chip, IEEE Spectrum, Jul. 1990.
`Wenzel, et al., A Multisensor Employing an Ultrasonic
`Lamb-Wave Oscillator, IEEE Transactions on Electron
`Devices, vol. 35, No. 5, Jun. 1988.
`Angell, et al., Silicon Micromechanical Devices, Scientific
`American 248:44-55, 1983.
`Petersen, Silicon as a Mechanical Material, Proceedings of
`the IEEE, vol. 79, No. 5, May 1982.
`
`THERMO FISHER EX. 1044
`
`
`

`

`U.S. Patent
`
`Dec. 17, 1996
`
`Sheet 1of10
`
`5,585,069
`
`16 7
`
`20
`
`18
`
`24125126
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`14
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`FIG. 1A
`
`FIG. 18
`
`THERMO FISHER EX. 1044
`
`
`

`

`U.S. Patent
`
`Dec. 17, 1996
`
`Sheet 2of10
`
`5,585,069
`
`14
`
`FIG. 2
`
`(25
`1
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`
`FIG. 3
`
`THERMO FISHER EX. 1044
`
`
`

`

`U.S. Patent
`
`Dec. 17, 1996
`
`Sheet 3of10
`
`5,585,069
`
`54
`
`55
`
`55"'-
`
`FIG. 4A
`
`38
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`56
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`

`

`U.S. Patent
`
`Dec. 17, 1996
`
`Sheet 4of10
`
`5,585,069
`
`48
`
`56
`
`FIG. 6A
`
`FIG. 68
`
`FIG. 6C
`
`THERMO FISHER EX. 1044
`
`
`

`

`U.S. Patent
`
`Dec. 17, 1996
`
`Sheet 5 of 10
`
`5,585,069
`
`16
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`

`U.S. Patent
`
`Dec. 17, 1996
`
`Sheet 7of10
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`5,585,069
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`
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`

`

`U.S. Patent
`
`Dec. 17, 1996
`
`Sheet 8 of 10
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`5,585,069
`
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`

`

`U.S. Patent
`
`Dec. 17, 1996
`
`Sheet 9of10
`
`5,585,069
`
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`
`
`

`

`U.S. Patent
`
`Dec. 17, 1996
`
`Sheet 10 of 10
`
`5,585,069
`
`442
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`
`THERMO FISHER EX. 1044
`
`
`

`

`5,585,069
`
`1
`PARTITIONED MICROELECTRONIC AND
`FLUIDIC DEVICE ARRAY FOR CLINICAL
`DIAGNOSTICS AND CHEMICAL SYNTHESIS
`
`This invention relates to a system comprising a parti-
`tioned microelectronic and fluidic array. More particularly,
`this invention relates to a system including an array of
`microelectronic and fluid transfer devices for carrying out
`various processes, including syntheses, screening and
`chemical diagnostic assays, in parallel, and method of 10
`making the array.
`
`5
`
`BACKGROUND OF THE DISCLOSURE
`
`15
`
`Traditional methods of making a homologous series of
`compounds, or the testing of new potential drug compounds
`comprising a series of like compounds, has been a slow
`process because each member of the series or each potential
`drug must be made individually and tested individually. For
`example, a plurality of potential drug compounds is tested 20
`by using an agent to test a plurality of materials that differ
`perhaps only by a single amino acid or nucleotide base, or
`have a different sequence of amino acids or nucleotides.
`Recently the process has been improved somewhat by
`combining the synthesis of various compounds having
`potential biological activity, for example, and traditional
`semiconductor techniques. A semiconductor or dielectric
`substrate for example is coated with a biologic precursor
`having amino groups with a light-sensitive protective chemi­
`cal attached thereto, and a series of masks are placed over
`the substrate, each mask having an opening. A coupling
`agent, such as a photosensitive amino acid, is illuminated
`through the opening, forming a particular compound by
`reaction with the amino compound. Additional masks are
`used with different coupling agents to form an array of 35
`different peptides on the substrate which array can then be
`tested for biologic activity. Suitably this is done by exposure
`of the array to a target molecule, such as an antibody or a
`virus. The array is exposed to a biologic receptor having a
`fluorescent tag, and the whole array is incubated with the 40
`receptor. If the receptor binds to any compound in the array,
`the site of the fluorescent tag can be detected optically. This
`fluorescence data can be transmitted to a computer which
`can compute which compounds reacted and the degree of
`reaction. This technique permits the synthesis and testing of 45
`thousands of compounds in days rather than in weeks or
`even months.
`However, the synthesis of each coupling reaction is not
`al ways complete, and the yield decreases as the length of the
`biopolymer increases. The process of aligning a plurality of
`masks and forming openings in the masks in sequence
`requires careful alignment and takes time.
`The above synthesis is made possible by two other recent
`technical developments that allow various manipulations
`and reactions on a planar surface. One is the detection and
`analysis of DNA fragments and their identification by reac­
`tion with specific compounds. Probes, RNA and DNA
`fragments can be resolved, labeled and detected by high
`sensitivity sensors. The presence or absence of DNA frag- 60
`ments can identify diseased cells for example.
`Another step forward is the ability to separate materials in
`a microchannel, and the ability to move fluids through such
`microchannels. This is made possible by use of various
`electro-kinetic processes such as electrophoresis or electro- 65
`osmosis. Fluids may be propelled through very small chan­
`nels by electro-osmotic forces. An electro-osmotic force is
`
`2
`built up in the channel via surface charge buildup by means
`of an external voltage that can "repCI" fluid and cause flow.
`This surface charge and external voltage produces an elec-
`tro-kinetic current that results in fluid flow along the chan­
`nel. Such electro-kinetic processes are the basis for a device
`described by Pace in U.S. Pat. No. 4,908,112 for example.
`Thus real progress has been made using electrophoresis
`and/or electro-osmosis to move very small amounts of
`materials along microchannels. Such movement can be used
`for synthesizing very small samples of potential drug com­
`pounds in an array and testing very small amounts of
`materials for bioactivity. Further progress in fully automat­
`ing the fiuidic processes will result in the synthesis and
`testing of vast numbers of compounds for bioactivity of all
`types, which information can be made available for future
`drug selection and will greatly reduce the time and expense
`of such testiJ::ig.
`
`SUMMARY OF THE INVENTION
`
`The system of the invention comprises a device array of
`micron sized wells and connecting channels in a substrate
`that interfaces with a station for dispensing fluids to and
`collecting fluids from, the array, and for performing electro-
`25 optic measurements of material in the wells. The station is
`also connected to control apparatus means to control the
`fluid flow to the channels and wells and to collect measure­
`ment data from the substrate. The above elements are
`interdependent and together can perform a variety of tasks in
`30 parallel.
`The individual wells of the array and their sequence can
`be varied depending on the synthesis or analysis to be
`performed. Thus the function of the arrays can be readily
`changed, with only the additional need to choose suitable
`interface means for monitoring and controlling the flow of
`fluids to the particular array being used and the test or
`synthesis to be performed.
`In one embodiment the above system can be used to
`perform various clinical diagnostics, such as assays for DNA
`in parallel, using the known protocols of the polymerase
`chain reaction (PCR), primers and probe technology for
`DNA assay. In another embodiment the above system can be
`used for immunoassays for antibodies or antigens in parallel
`for screening purposes. In still other embodiments, the
`synthesis of a series of chemical compounds, or a series of
`peptides or oligonucleotides, can be performed in parallel.
`Each well in the array is designed so to accomplish a
`selected task in appropriate modules on a substrate, each
`50 module containing the number of wells required to complete
`each task. The wells are connected to each other, to a sample
`source and to a source of reagent fluids by means of
`connecting microchannels. This capability permits broad
`based clinical assays for disease not possible by sequential
`assay, permits improvement in statistics of broad based
`clinical assays such as screening of antibodies because of the
`parallelism, permits a reduction in costs and an improve­
`ment in the speed of testing, and permits improved patient
`treatments for rapidly advancing disease.
`The array is formed in a suitable dielectric substrate and
`the channels and wells are formed therein using maskless
`semiconductor patterning techniques. The station and con­
`trol means, such as a computer, use existing technology that
`includes commercially available apparatus.
`The present device array uses active control to move
`fluids across the array, reducing the time required for syn­
`thesis and screening. Further, large biopolymers of all types
`
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`can be synthesized and processed while maintaining high
`purity of the synthesized compounds. The present micro­
`laboratory arrays may be fully automated, enabling the rapid
`transfer of samples, precursors and other movement of fluids
`into the array, from one well to another well, and to enable
`the measurement of assays and the complete control of
`processing parameters such as temperature control.
`
`BRIEF DESCRIPTION OF THE DRAWING
`
`5
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`10
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`15
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`20
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`4
`The system of FIG. lA includes a computer 10, electri­
`cally connected via line 11 to peripheral apparatus, such as
`a modem or printer 12, which computer 10 is programmed
`to give instructions to a microlaboratory disc 14 and to
`record test results obtained therefrom. The computer 10 is
`electrically connected to a station 16 via line 15. The station
`16 includes a microlaboratory disc support 18, support
`tubing 20 for loading test materials and reagents onto the
`microlaboratory disc 14, pumps 22 for moving fluids to
`particular destinations on the disk 14, one or more light
`sources 24, an optical fiber 25 and one or more light
`detectors 26. The optical fiber 25 is operative to transmit
`light from the light source 24 to the detector 26. One or more
`containers 28 for waste fluids and the like are also housed in
`the station 16.
`A small volume of a fluid to be tested, such as a whole
`blood or other DNA-containing fluid sample, is loaded into
`the system via a loading system 30. The system 30 may
`house one or more capillary tubes 32, illustrated in FIG. 3,
`containing a sample which is connected to a loading capil-
`lary channel 34 etched into the surface of the rnicrolabora­
`tory disc 14. The loading capillary tube 32 is inserted into
`the capillary loading channel 34 horizontally. The sample is
`moved into the capillary loading channel 34 and the fluid
`sample is then moved to a first well 36 on the disc 14.
`Alternately, a loading channel 50 for vertical insertion into
`the loading channel 34 can also be used to load the sample.
`FIG. lB and FIG. 2 illustrate a particular module comprising
`a plurality of wells 36, 40, 42 and 44 connected by a channel
`38, shown by a dashed line, on a rnicrolaboratory disc 14 of
`the invention. Each module is connected to a well 46 that
`collects excess or waste fluids from the respective module.
`These waste fluids are collected and moved to the waste
`containers 28 in the station 16.
`The sample is treated sequentially in a series of wells
`including first well 36. For example, in the first well 36 the
`whole blood sample is transferred from the capillary loading
`channel 34, filtered and lysed to separate the white and red
`corpuscles, and the DNA is isolated from the white blood
`40 cells. The DNA sample is then moved out of the first well 36
`through a connecting channel 38 that connects all of the
`wells of a single module, and into a second well 40. In the
`second well 40 the DNA is separated into single strands and
`amplified using the well known PCR method. The treated
`sample is then moved out of the second well 40 via the
`connecting channel 38 and into a third well 42. In the third
`well 42 the DNA is assayed by known probe hybridization
`techniques. The DNA assay is detected and evaluated in the
`fourth well 44. Thus the determination of DNA in a par-
`ticular blood sample is performed in a series of four wells
`connected by a channel. Lastly excess reagents and the like
`are collected in the fifth well 46 that is common to all of the
`modules in the rnicrolaboratory array 14, and is transferred
`into the waste collection system 28 of the station 16.
`The combination of a loading channel 34 or 50, the wells
`36, 40, 42, 44 and 46 and the connecting channel 38 make
`up one module 48 on the test microlaboratory disc 14. A
`single module on a microlaboratory disc 14 is shown in top
`view in FIG. lB within the dashed lines. As will be
`60 explained hereinbelow, a plurality of modules are formed in
`the rnicrolaboratory disc 14 so that tests can be performed on
`a large number of the modules 48 in parallel.
`The particular sequence of channels and wells of the
`module shown in FIGS. lA, 1B and FIG. 2 lends itself to the
`detection of pathogenic bacteria in blood or other DNA­
`containing fluids using the DNA assay protocols of Greisen
`et al, see "PCR Primers and Probes for the 16S rRNA Gene
`
`FIG. lA is an exploded schematic diagram of the parts of
`the system of the invention adapted for performing clinical
`assays.
`FIG. lB is a top view of a substrate of the invention
`illustrating a single module formed therein.
`FIG. 2 is an exploded top view of an illustrative module
`of the invention.
`FIG. 3 is an exploded cross sectional view of the optical
`transmission and detection system of the invention.
`FIG. 4A is a cross sectional view of a well embodiment
`of a microlaboratory disc of the invention.
`FIG. 4B is a cross sectional view of another well embodi­
`ment of a rnicrolaboratory disc of the invention.
`FIG. SA is a cross sectional view of a portion of a module 25
`of a microlaboratory disc illustrating devices in typical
`wells.
`FIG. SB is a cross sectional view of a portion of a module
`of a microlaboratory disc covered with a cover plate.
`FIG. 6A is a cross sectional view of a rnicrolaboratory
`disc illustrating additional wells having preformed devices
`therein together with an optical system interface.
`FIGS. 6B and 6C illustrate a valve situate in a channel
`adjacent to a well in the open and closed positions respec­
`tively.
`FIG. 7 A is an exploded schematic view of another
`embodiment of the present invention adapted to perform
`immunological assays.
`FIG. 7B is a top view illustrating a module on the
`microlaboratory disc of FIG. 7 A.
`FIG. 7C is a cross sectional view of a control means for
`moving fluids in the channels and from one well to another.
`FIG. 8 is a schematic view of another embodiment of a 45
`rnicrolaboratory array suitable for carrying out the parallel
`synthesis of proteins and oligonucleotides.
`FIG. 9 is a schematic view illustrating a modified station
`for the system of FIG. 8.
`FIG. 10 is a schematic view of a further embodiment of 50
`a rnicrolaboratory array suitable for carrying out the syn­
`thesis of a large number of small molecules in parallel.
`FIGS. llA, llB and UC are cross sectional views illus­
`trating the steps needed to form cross over channels in the
`substrate.
`FIG. llD is a top view of a cross-over channel in the
`substrate.
`FIG. 12 is a top view of a channel "gate" electrode to
`control flow in a channel by electro-osmosis.
`
`30
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`35
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`DETAILED DESCRIPTION OF THE
`INVENTION
`
`The invention will be first described by reference to FIG. 65
`lA, which illustrates the parts of an illustrative system of the
`invention configured to perform DNA screening diagnostics.
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`of Most Species of Pathogenic Bacteria, Including Bacteria
`Found in Cerebrospinal Fluid" J. Clinical Microbiology 32
`(2), pp 335-351 (1994). Since the disc 14 can be arranged
`to form a plurality of up to 1500 parallel modules, broad
`DNA screening can be conducted in parallel that might not 5
`be possible using a system of sequential assays. Since a large
`array of parallel tests can be done at the same time, the time
`required to obtain meaningful results is also reduced, and
`consequently costs are reduced. Further, the statistics of
`broad based screening can be improved. In addition the
`system of the invention provides the ability to detect mutant
`DNA such as is found in oncological diseases.
`The station 16 includes a fiber optic assembly 25 and one
`or more light sources 24 and one or more detectors 26 (not
`shown) that address the system loading channel 34 or 50 and
`measures the transmittance or absorbance of material in the 15
`channel 34 or 50 and the first well 36, such as a blood or
`other fluid sample. The fiber optic assembly 25 can verify
`the presence or absence of materials in the channel 34 or 50
`or the well 36, and quantify their amounts by transmitting
`the measurement data to the computer 10. Suitable lasers 20
`and photodetectors are available commercially. Fiber optic
`adaptors to support the optical fiber are commercially avail­
`able. These adaptors may also include a lens for efficient
`transfer of light from the light source into the fiber.
`Preselected substrates, designed either for modular par-
`allel or array processing, can be placed on the substrate
`holder 18 in the station 16. After suitable modification of the
`software in the computer 10 or choice of other driving means
`able to monitor a particular sequence of steps for control of
`reagents to carry out the processes in each module of the
`microlaboratory 14, entirely different tests and processes can
`be carried out in the system of the invention. Thus the
`microlaboratory array 14 is individually designed for a
`variety of assay, measurement and synthesis tasks, and only 35
`the module configuration and the driving means needs to be
`changed in the system when a different task is to be
`performed.
`A circuit of thin film transistors (not shown) can be
`formed on the front or back of the glass or other substrate 14
`to provide power to the wells via leads and electrodes
`explained further hereinafter, and to connect them with the
`driving means such as the computer 10, so as to move liquids
`along the array. Pins can also be formed in the glass substrate
`which are addressable by logic circuits on the support 18 that
`are connected to the computer 10 for example. These
`transistors and logic circuits will also be in contact with the
`substrate 14 on the support 18 to provide an interface
`between the microlaboratory disc 14 and the computer 10.
`The system loader 30 is situated outside of the station 16
`and may be connected to a loading capillary tube 32, as
`shown in FIG. 3, suitably having an inner diameter of about
`200 microns and an outer diameter of about 600-700
`microns. The capillary tube 32 can be pretreated to eliminate
`surface adsorption of proteins and related bio materials in a
`known manner, for example similar to the methods disclosed
`by Cobb, ''Electrophoretic Separations of Proteins in Cap­
`illaries with Hydrolytically Stable Surface Structures", Anal.
`Chem. 62, (1990) pp 2478-2483. All of the sample material
`is loaded into the system via the system loader 30. The
`system capillary tube 32 can be loaded horizontally directly
`into the microlaboratory disk 14 via the loading channel 34.
`The sample is i