EP0892672A1 - Plaque pour systeme de reaction - Google Patents

Plaque pour systeme de reaction

Info

Publication number
EP0892672A1
EP0892672A1 EP97917901A EP97917901A EP0892672A1 EP 0892672 A1 EP0892672 A1 EP 0892672A1 EP 97917901 A EP97917901 A EP 97917901A EP 97917901 A EP97917901 A EP 97917901A EP 0892672 A1 EP0892672 A1 EP 0892672A1
Authority
EP
European Patent Office
Prior art keywords
cells
plate
gasket
cell
reaction
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP97917901A
Other languages
German (de)
English (en)
Other versions
EP0892672A4 (fr
Inventor
Robert Richard Demers
Satyam Choudary Cherukuri
Aaron William Levine
Kerry D. O'mara
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Sarnoff Corp
Original Assignee
Sarnoff Corp
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from US08/630,018 external-priority patent/US5840256A/en
Priority claimed from US08/744,386 external-priority patent/US6033544A/en
Application filed by Sarnoff Corp filed Critical Sarnoff Corp
Publication of EP0892672A1 publication Critical patent/EP0892672A1/fr
Publication of EP0892672A4 publication Critical patent/EP0892672A4/fr
Withdrawn legal-status Critical Current

Links

Classifications

    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5025—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures for parallel transport of multiple samples
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
    • B01F35/00—Accessories for mixers; Auxiliary operations or auxiliary devices; Parts or details of general application
    • B01F35/71—Feed mechanisms
    • B01F35/717—Feed mechanisms characterised by the means for feeding the components to the mixer
    • B01F35/7182—Feed mechanisms characterised by the means for feeding the components to the mixer with means for feeding the material with a fractal or tree-type distribution in a surface
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J19/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J19/0046—Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J19/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J19/0093—Microreactors, e.g. miniaturised or microfabricated reactors
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
    • B01F25/00—Flow mixers; Mixers for falling materials, e.g. solid particles
    • B01F25/40—Static mixers
    • B01F25/41—Mixers of the fractal type
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00277—Apparatus
    • B01J2219/00279—Features relating to reactor vessels
    • B01J2219/00306—Reactor vessels in a multiple arrangement
    • B01J2219/00313—Reactor vessels in a multiple arrangement the reactor vessels being formed by arrays of wells in blocks
    • B01J2219/00315—Microtiter plates
    • B01J2219/00317—Microwell devices, i.e. having large numbers of wells
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00277—Apparatus
    • B01J2219/00351—Means for dispensing and evacuation of reagents
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00583—Features relative to the processes being carried out
    • B01J2219/00585—Parallel processes
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00583—Features relative to the processes being carried out
    • B01J2219/0059—Sequential processes
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00583—Features relative to the processes being carried out
    • B01J2219/00596—Solid-phase processes
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00781—Aspects relating to microreactors
    • B01J2219/00819—Materials of construction
    • B01J2219/00824—Ceramic
    • B01J2219/00828—Silicon wafers or plates
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00781—Aspects relating to microreactors
    • B01J2219/00819—Materials of construction
    • B01J2219/00831—Glass
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00781—Aspects relating to microreactors
    • B01J2219/00851—Additional features
    • B01J2219/00853—Employing electrode arrangements
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00781—Aspects relating to microreactors
    • B01J2219/00891—Feeding or evacuation
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00781—Aspects relating to microreactors
    • B01J2219/0095—Control aspects
    • B01J2219/00952—Sensing operations
    • C—CHEMISTRY; METALLURGY
    • C40—COMBINATORIAL TECHNOLOGY
    • C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
    • C40B60/00—Apparatus specially adapted for use in combinatorial chemistry or with libraries
    • C40B60/14—Apparatus specially adapted for use in combinatorial chemistry or with libraries for creating libraries

Definitions

  • the present invention relates to a plate having formed thereon numerous small- scaled reaction cells (or wells) that are formatted to allow the cells to be individually addressed despite their small size, to allow each cell to accommodate a substrate for reactions, to allow each cell to be separately illuminated for optical detection, and to allow sufficient material between reaction cells to support the formation of an appropriate seal between the plate and an ancillary device, for instance a liquid distribution device that delivers reagents to the reaction cells.
  • an ancillary device for instance a liquid distribution device that delivers reagents to the reaction cells.
  • Such devices allow the programming of the concurrent operation of thousands of separate reactions.
  • Pursuant to the Zanzucchi et al. patent application such an apparatus can be used to distribute various liquids to thousands of reaction cells, which reaction cells can be fabricated on a plate.
  • microtiter plates having 96 or 384 cells to assay prospective pharmaceuticals in an in vitro model for a pharmaceutical activity or for conducting small-scale syntheses in parallel.
  • the devices provided for by the advances in microfluidics are to be used to conduct reactions at, for example, 1,000, 10,0000, 100,000 or more reaction sites or cells. By the present invention, these sites or cells are located on a plate.
  • the invention provides a plate having a plurality uniformly sized reaction cells formed in its upper surface, wherein the density of the reaction cells is at least about 10 cells per cm ⁇ .
  • the reaction cells are arrayed in rows and columns.
  • the plate is rectangular, preferably with the rows and columns of cells parallel to the edges of the plate.
  • the area of each of the openings (i.e., apertures) of the reaction cells is no more than about 55% of the area defined by the multiplication product of (1) the pitch between reaction cells in separate rows and (2) the pitch between reaction cells in separate columns.
  • the product between this row pitch and column pitch can be termed the "footprint" of a cell, meaning the amount of area supporting each cell of a plate.
  • this aperture area is no more than about 50%, yet more preferably 45%, of the cell footprint.
  • the density of cells is no more than about 350 per cm ⁇ , more preferably no more than about 150 per cm ⁇ , yet more preferably no more than about 120 per cn v
  • the density of cells is at least about 10 cells per cm ⁇ , more preferably at least about 20 cells per crn ⁇ , more preferably at least about
  • the footprint which is symmetrically overlaid on the cell, encompasses plate surface area on all sides of a cell.
  • the minimum distance from the edge of the cell to the boundary of the footprint is here termed the "seal strip width," since this is the area on which a gasket material can be applied.
  • the density of cells is from about 10 to about 45 cells per cm 2 , more preferably about 10 to about 20 cells per cm 2
  • the seal strip width is from about 300 ⁇ m to about 1,000 ⁇ m, more preferably from about 600 ⁇ m to about 1,000 ⁇ m.
  • the diameter or width of the aperture of a cell is about 400 ⁇ m to about 1 100 ⁇ m, more preferably about 900 ⁇ m to about 1 100 ⁇ m.
  • the depth of the cells is preferably from about 100 ⁇ m to about 400 ⁇ m, more preferably about 250 ⁇ m to about 350 ⁇ m
  • the pitch between reaction cells in a row or column is at least about 0 5 mm, more preferably at least about 0 9 mm
  • each reaction cell is separated from each adjacent reaction cell by at least about 0 15 mm, more preferably by at least about 0 3 mm
  • each reaction cell has a substantially square shape
  • the plate has at least about 1,000 reaction cells, more preferably at least about 4,000 reaction cells, yet more preferably at least about 10,000 reaction cells
  • the plate has a patterned gasket on its upper surface
  • the plate is designed to facilitate alignment by having a first marker on a first edge of the plate, wherein the marker is for orienting the reaction cells
  • the plate has a second marker on a second edge of the plate perpendicular to the first edge, wherein the second marker is for orienting the reaction cells
  • the plate has a third marker on the second edge, wherein the third marker is for orienting the of reaction cells
  • the first marker on a first edge of the plate
  • the plate has a second marker on a second
  • the invention also provides a reaction system for conducting a plurality of reactions in parallel, the reaction system comprising a liquid distribution system for addressably directing a plurality of liquids to a plurality of cells, and a plate as described above
  • the invention additionally provides a method of conducting a plurality of reactions in parallel comprising operating a liquid distribution system for addressably directing a plurality of liquids to a plurality of cells, wherein the cells are located on one of the above- described plates that is reversibly sealed to the liquid distribution system
  • the invention further provides a method of forming a gasket on the top surface of a substrate, the method comp ⁇ sing patterning a layer of photoresist on the top surface so that there are cleared surface areas and photoresist-coated areas, applying an elastomeric gasket material to the cleared areas, and removing the photoresist from the photoresist-coated areas
  • the applying step comprises placing the elastomeric gasket material on the top surface, and compression-molding the gasket material into the cleared areas
  • the gasket material is cured
  • the gasket material is preferably silicone
  • the substrate preferably has a smooth surface, more preferably a flat surface
  • the method is preferably applied to a plate on which structures, particularly microstructures, have been formed BRIEF DESCRIPTION OF THE DRAWINGS
  • Figure 1 depicts four formats for the plate of the invention
  • Figure 2 shows schematically some of the parameters considered in designing the format of the plate of the invention
  • Figures 3A-3C show three cell designs
  • Figure 4 illustrates a plate of the invention connected to a liquid distribution system
  • Figure 5 shows a plate of the invention
  • Figure 6A and 6B illustrate a portion of a gasket print pattern DEFINITIONS
  • addressable Cells are addressable if each cell of a plate can be individually located and its contents manipulated
  • “Alignment” refers to the coordinated positioning of the cells of a small-scaled plate of the invention and another device with which the small-scaled plate will operate
  • the small-scaled plate is aligned with a liquid distribution system if the liquid distribution system is positioned to deliver liquid to each of the cells of the small-scaled plate
  • the small-scaled plate is aligned with an optical detection device if the device can focus light on each cell and separately identify the transmission or fluorescence of each cell
  • the concept relates to the relative positioning between a device, such as a fabrication device, and the design location of the cells of the small-scaled plate as set forth in the design plans • micromachining
  • Micromachining is any process designed to form microstructures on a substrate. structure
  • a “structure” is formed on the upper surface of a plate is a shape defined by variations in the elevation of the upper surface
  • structures are “microstructures” having dimensions of about 2 mm or less
  • the small-scaled plate 100 of the invention is formed of a substrate that is an organic or inorganic material that is suitable for forming the fine structures described herein
  • the small-scaled plate 100 should be formed of a material that is resistant to the types of materials it is anticipated to encounter in use Thus, for instance, in diagnostic settings the small-scaled plate 100 typically encounters aqueous materials and can, accordingly, be manufactured of a broad range of materials Where the small-scaled plate
  • the small-scaled plate 100 is designed for use in synthetic reactions, often the 100 should be constructed of a material that is resistant to acids, bases and solvents
  • the small-scaled plate 100 is constructed of glass, particularly borosilicate glass
  • a basic parameter for the small-scaled plate 100 is the spacing between the centers of adjacent cells 101 , which spacing is termed the "pitch"
  • Four cell formats for plates are illustrated in Figure 1 , these formats are the IK, 4K, 10K and 100K formats
  • the IK format has a pitch of 2260 ⁇ m; the 4K format has a pitch of 1488 ⁇ m, the 10K format has a pitch of 965 ⁇ m, and the 100K format has a pitch of 558 ⁇ m
  • Illustrative parameters for these formats are set forth below
  • cell volume and depth are selected to help accommodate the insertion of beads on which synthetic or other chemistries are conducted.
  • the pitch is the 2260 ⁇ m distance illustrated in Figure 1.
  • the area defined by the pitch further defines the amount of surface area that a given cell 101 resides within.
  • the product of the pitch between cells 101 in a row and the pitch between cells 101 in a column determines the size of the surface area on which an individual cell 101 sits
  • the percentage of this surface area taken up by the area of each of the cell apertures is the area of the cell openings divided by the above-described product, times 100% It is useful in understanding how the small-scaled plate is used, to refer to
  • the reaction cells are preferably found on a plate 100 that is separable from the portion of the liquid distribution system containing reservoirs and pumps
  • the separable plate 100 docks with the liquid distribution system, typically with a gasket material (that has openings at appropriate locations) interposed between the two, so that the cells are aligned underneath the appropriate outlet for delivering liquid from the liquid distribution system.
  • Three parameters that are basic to the format of the plate 100 are the spacing between cells 101 (i e., pitch), the area of each of the openings of the cells 101 which will be referred to as the cell aperture, and the row-column arrangement which will be referred to as the matrix layout
  • the depth of a cell 101 can be made to vary according to the application for which the plate 100 is used. Structures required for support functions can be formed on the area between cell apertures
  • the determination of pitch is based on factors including the following:
  • the determination of cell aperture is based on factors including the following:
  • the determination of the matrix layout is based on factors including the following:
  • Such Designs are illustrated in Figure 1.
  • Intermediate formats covering a different number of cells 101, and asymmetric matrix layouts can also be fabricated.
  • Format 1 K is a 1024 cell array symmetrically formed into 32 rows and 32 columns and having a reaction cell volume of at least about 120 nanoliter per cell 101.
  • the substrate size has been selected by balancing the pressures toward minimum size lmposed by handling and fluidics factors and the pressures for maximum size imposed by ease of fabrication considerations Using approaches that most reflect the performance implied by this format, a substrate size of 3 inch x 3 inch has been selected as the best compromise
  • a cell pitch of 2260 ⁇ m can be accommodated
  • a cell configuration that satisfies volumetric and surface area requirements for fluid delivery, synthesis, assay and detection is 890 ⁇ m x 890 ⁇ m
  • the cells 101 have a fluid capacity of a minimum of 120 nano ters Format 4K
  • Format 4K is a 4096 cell array symmetrically formed into 64 rows and 64 columns and having a reaction cell volume capacity of at least about 120 nanoliter per cell 101
  • a compromise between operation, handling and fabrication has led to the selection of a substrate size of 4 inch x 4 inch
  • a cell pitch of 1488 ⁇ m can be accommodated
  • the cell configuration of 890 ⁇ m square of the IK format which configuration satisfies volumetric and surface area requirements for fluid delivery, synthesis, assay, and detection, can be maintained Using typical micromachining techniques suitable for production, the cells 101 have a fluid capacity of a minimum of 120 nanohters Format 10K
  • Format 1 OK is a 10,000 cell array symmetrically formed into 100 rows and 100 columns A maximum of 4 inch x 4 inch substrate size was selected for handling and fabrication reasons Micromachined features are reduced in size from the 4K cell format
  • the associated liquid distribution system for instance a liquid distribution system according to Zanzucchi et al .”Liquid Distribution System," U S Patent Application No 08/556,036, filed November 9, 1995, is also fabricated with a correspondingly dense layout of fluid delivery capillaries With such a dense layout of fluid delivery capillaries, a cell pitch of 965 ⁇ m in the small-scaled plate can be accommodated
  • the cell configuration is adjusted for the more demanding requirements created by the higher density of cells
  • a useful resolution of the volumetric and surface area requirements for fluid delivery, synthesis, assay and detection is 635 ⁇ m x 635 ⁇ m cell aperture.
  • the cells 101 have a fluid capacity of a minimum of 50 nanohters.
  • the reaction cell aperture is preferably substantially square or rectangular in profile to best accommodate an array format.
  • the aperture can have rounded corners to accommodate the micromachining or molding/replication techniques used.
  • substantially in this context means no more than the amount of rounding or irregularity in shape that can be expected when such structures are formed in glass by chemical etching, as predominately practiced commercially in 1995.
  • the circular features formed at the edges of the "rectangular” or “square” cells have radii no greater than the depth of the cell and the edges of the aperture of the cell are longer than the cell depth.
  • first cell 101 A illustrates the relatively sharp edge lines obtained by chemically etching a silicon substrate.
  • second cell 101B illustrates the relatively sharp edge lines obtained by laser etching a glass substrate.
  • the lines obtained are typically less sharp, as illustrated in Figure 3C.
  • the cell 101 cross-sectional profile can be of various shapes depending on the micromachining or replication technique but should preferably meet a minimum fluid volume capacity and must provide enough depth to accommodate experiments that require a bead 102 for use in syntheses or assays that require a solid support. Although a number of beads per cell may be used, and although beads of different sizes may be used depending on the experiment, the preferred design consideration is based on providing adequate space for synthesis or other reaction on a single bead of a defined maximum specified swollen diameter.
  • cell depths sufficient to accommodate swollen beads of about 200 ⁇ m diameter, or even about 400 ⁇ m, are used in formats IK, 4K; and depths sufficient to accommodate swollen beads of 100 ⁇ m diameter are used in format 100K.
  • the cell profile is achieved with micromachining, replicating, molding, or like fabrication methods, cells in a single substrate, or is achieved by combining multiple layers of substrates.
  • the combining of layers can be achieved by known methods or, with approp ⁇ ate substrates, with the field-assist sealing method described in Zhonghui H Fan et al , U S Provisional Application No P-89,876, titled "Field Assisted Glass-Glass Sealing,” filed November 7, 1995, which is incorporated herein in its entirety by reference
  • the small-scaled plate allows for the space between cells to be used to provide for fluid conduits and drains, electrical vias, sealing features, and the like
  • the small-scaled plate can be constructed of any materials, material combinations, substrate thicknesses, and fab ⁇ cation techniques, that suit the application Figure 4 illustrates one way the surface area between cell apertures can be used
  • the small-scaled plate 320 is formed of a single plate and has formed thereon cells 350
  • the small-scaled plate 320 is designed for use with a liquid distribution system ("LDS") formed of first LDS plate 300 and second LDS 310 Liquid is delivered to each cell 350 through a first conduit 390 Excess fluid flows out through second conduit 355, which connects to cell 350 through third conduit 351
  • LDS liquid distribution system
  • mechanical alignment using three-pin registry is acceptable, and the edge alignment locations specified in Figure 5 can be used
  • the preferred method is to grind first edge notch 105 A, second edge notch 105B and third edge notch 105C, for instance at the locations shown in Figure 5
  • the use of such notches obviates the need to accurately machine all the edges of the small-scaled plate 100 and provides for a method of mechanically identifying the top and bottom of the plate 100
  • the location of the center of the cell patterns is defined in Figure 5 by the intersection of lines B and C
  • the use of comparable notches in the manufacture of a liquid distribution system with which the small-scaled plate operates allows equipment and tool manufacturers to coordinate their designs
  • examples of the distances represented by Rl , R2, R3, Cm and Co are FORMAT Rl R2 R3 Cm Co
  • the preferred support material will be one that has shown itself susceptible to microfab ⁇ cation methods, such as a microfab ⁇ cation method that can form channels having cross-sectional dimensions between about 50 microns and about 250 microns
  • Such support materials include glass, fused silica, quartz, silicon wafer or suitable plastics Glass, quartz, silicon and plastic support materials are preferably surface treated with a suitable treatment reagent such as a siliconizing agent, which minimizes the reactive sites on the material, including reactive sites that bind to biological molecules such as proteins or nucleic acids
  • a non-conducting support material such as a suitable glass
  • Preferred glasses include borosilicate glasses, low-alkali lime-silica glasses, vitreous silica
  • reaction cells and horizontal channels and other structures of the small-scaled plates can be made by the following procedure
  • a plate is coated sequentially on both sides with, first, a thin chromium layer of about 50 ⁇ A thickness and, second, a gold film about 2000 angstroms thick in known manner, as by evaporation or sputtering, to protect the plate from subsequent etchants
  • a two micron layer of a photoresist such as Dynakem EPA of Hoechst-Celanese Corp , Bndgewater, NJ, is spun on and the photoresist is exposed, either using a mask or using square or rectangular images, suitably using the MRS 4500 panel stepper available from MRS Technology, Inc., Acton, MA.
  • the gold layer in the openings is etched away using a standard etch of 4 grams of potassium iodide and 1 gram of iodine (I2) in 25 ml of water.
  • the underlying chromium layer is then separately etched using an acid chromium etch, such as KTI
  • the gasket used to reversibly seal the plate to an instrument that functions with the plate can be attached to the plate, leaving openings for the cells and other structures, as needed.
  • One method of attaching the gasket is screen-printing.
  • the printed gasket can be made of silicone or another chemically-resistant, resilient material.
  • a multi-step compression-molding process that utilizes photolithography can be applied.
  • the top surface of the plate, on which generally cells and other structures have been formed is coated with a photoresist.
  • the photoresist layer is about 1 mil in thickness
  • the photoresist layer is treated by standard photolithography techniques to remove photoresist from those areas (the "gasket areas") away from the apertures of the cells where gasket material is desired.
  • a layer of a flowable gasket material that can be cured to a resilient, elastomeric solid is applied.
  • a platen having a polished surface, for instance a polished glass surface is placed above the gasket material and pressure is applied to push the gasket material into the gasket areas and substantially clear the gasket material from the photoresist-coated areas.
  • the gasket material is now cured
  • the photoresist is then dissolved, leaving the plate with a patterned gasket.
  • the gasket material is substantially cleared if it is sufficiently cleared to allow the underlying photoresist to be dissolved.
  • the gasket material is any elastomeric material that is suitable for use in the above-described compression molding technique, that is, when cured, compatible with the chemistries that are to be practiced in the plate on which the gasket is formed, and that is, when cured, resistant to the solvents used to remove the photoresist.
  • the gasket material is preferably silicone, such as RTV type silicone rubber (e.g., Silastic J, RTV Silicone Rubber available from Dow Corning, Midland, Michigan).
  • the photoresist can be a film-type photoresist such that typically the structures on the plate will not be filled during the compression-molding process or a liquid-type photoresist such that the structures will temporarily be filled during the compression-molding process and etched away at the completion of the process.
  • a primer for promoting the adhesion of the gasket material such as 1200 RTV Prime Coat from Dow Corning, Midland, Michigan.
  • the plate can also be roughened to promote the adhesion of the gasket material to the plate. For example, 5 micron roughness can be produced by lapping.
  • the platen is preferably treated with a release-promoter, or a release promoter is incorporated into the gasket material, as it is in Silastic J silicone rubber.
  • the compression-molding process can leave thin residues of gasket material at unwanted locations. These residues are laser cut away from the plate or, in some cases, are removed using a timed exposure to a solvent that dissolves the thin film of exposed gasket material residue without having substantial effect on the thicker layer of gasket material found at desired locations.
  • the printed gasket can be made of silicone or another chemically-resistant, resilient material.
  • the gasket is made of a mixture of (a) a silicone rubber-forming material such as that available
  • Sylgard 184 and MDX4-4210 are sold in two components. One component is an emulsion containing particles of silcone rubber and a polymerization catalyst and the second component is a preparation of a bi ⁇ valent monomer, which monomer serves to crosslink and thereby cure the silicone rubber.
  • Component one of MDX4-4210, i.e. the "elastomer component,” is made up of dimethylsiloxane polymer, reinforcing silica, and a platinum catalyst.
  • Component two of MDX4-4210 also contains dimethylsiloxane polymer, in addition to a polymerization inhibitor, and a siloxane crosslinker.
  • the components are generally mixed according to the manufacturer's recommendations. For example, for MDX4-4210, ten parts by weight of emulsion, i.e. elastomer, are mixed with one part of monomer solution, i.e curing agent.
  • inert fillers about 7.5% by weight of M-5 grade Cab-o- sil can be added to the Sylgard 184, or about 2-3% by weight of M-5 grade Cab-o-sil can be added to the MDX4-4210.
  • Filler can serve to thicken the pre-polymerized composition to improve its screen printing properties.
  • Gasket materials can generally be cured at room temperature, or curing, can be accelerated with, for example, heat. Prior to curing, the gasket-forming material is capable of flow, though generally viscous flow, which flow is sufficient to facilitate the screen printing process The gasket-forming material is also sufficiently adhesive to adhere either to the plate to which it will be applied or to an underlying first layer of gasket material
  • a first layer of gasket material is printed onto the plate and then cured.
  • a second layer of gasket material is overlaid on the first, a smooth platen of appropriate shape (generally very flat) is overlaid upon the printed gasket material so that a uniform weight is applied to the printed gasket material (while taking precautions to prevent destructive adhesions of gasket material to the platen such as described further below), and the gasket material is cured.
  • the use of two printings of gasket-forming material helps form a foundation of gasket material prior to the smoothing process effected after the second printing and to achieve the needed smoothness and uniform thickness of the sealing surface of the gasket.
  • This pressure should be selected to be, for the particular gasket-applying process, sufficiently high to create the needed uniformity during the curing process, but not so high as to overly compress cured portions of gasket material such that upon release of the pressure these portions re-expand and create a non-uniform seal thickness.
  • a si le print process can also be used, and such a single print process is generally preferre ⁇ since it is simpler and more readily applied to a production process.
  • FIG. 6B shows an illustrative print screen pattern, wherein gasket material is apphed between the closely spaced (here, for example 6 mils) lines After printing and processing, the applied gasket patterns are broadened For example, in applications using the two-p ⁇ nt process and 6 mil wide pattern on the p ⁇ nt screen, an 18 mils wide pattern has been produced
  • the fifty figure eight patterns outlined in dark lines represent the gasket about one hundred reaction wells on a reaction cell plate, with the individual wells (not shown) located within the two openings in the illustrated figure eight patterns
  • Figure 6B is illustrated a print screen pattern used to generate one of the figure eight patterns
  • the plate in this case a 2 X 2 inch glass plate, is cleaned in a Class 10,000 or, preferably, cleaner cleanroom environment
  • the plate is inspected under a microscope for lint and deposits These are removed with tweezers and by a stream of propanol or other solvent
  • the plate is wiped with a lint-free cloth and vapor cleaned with ethanol
  • the plate can be stored in a container
  • the gasket forming material is prepared by degassing the material (e g , MDX 4-4210 ) under vacuum Care is taken to align the register between the plate and the print screen
  • the plate can be aligned with three-pin registry with the notches indicated at the edges of the plate illustrated m Figure 6A
  • the gasket pattern is then p ⁇ nted on the plate, the plate is isolated in a clean container, and the gasket is cured by placing the container in a 70 C oven for 4 hours Then, the same gasket pattern is overlaid on the first A thin, preferably transparent, plastic film
  • a clean, smooth plate can be placed on top of the gasket, and a clamping pressure of, for example, 20 lbs. is applied to the gasket pattern of Figure 6 A.
  • a contacting interface for each seal segment should be visible.
  • the gasket should be stored in contact with a release film.
  • Illustrative single-print protocol The plate is prepared as described above, and a gasket is printed on the plate as described above, taking special care that the gasket- forming material is evenly applied by the print screen.
  • a transparent, plastic film which is coated with a release coating, is layered over the printed pattern, and a smooth, flat platen is positioned over the film and the underlying printed pattern.
  • the platen is impressed upon the pattern until it is met by mechanical stops that hold the platen at a uniform height above the plate.
  • the gasket is then cured while the platen is in place
  • the gasket can be tested and stored as described above.
  • the screen used in the printing can be formed for example using conventional photolithographic means.
  • the screen is for example woven from 0.9 mil stainless steel wire.
  • the weave pattern is preferably oriented at about a 45 degree angle from the printing (squeegee) direction.
  • the photolithographic emulsion on which the pattern is made in can be, for example, 2.5 mils thick With the 6 mil screen pattern width illustrated in Figure 6B and the 2.5 mil screen pattern depth, the gasket width after curing is typically about 18 mil and the thickness of the seal is typically about 1.3 mils In printings using MDX 4-4210, a typical product gasket has a hardness of about 65 durometer.
  • the width and thickness of the gasket can be varied by, for example, varying the dimensions of the screen pattern, varying the size of emulsion particles in the polymeric component of the gasket-forming material, varying the weight applied in the curing process, and adding additives to the gasket-forming material such as an inert filler.
  • the gasket-forming process while preferably applied to the flat plates contemplated in the preferred embodiments of the liquid distribution system, can also be applied to any other surfaces to which a complementary surface can be sealed via the gasket. Generally, such other surfaces will be sufficiently smooth so as to facilitate printing of the gasket and sealing to the complementary surface. While this invention has been described with an emphasis upon preferred embodiments, it will be obvious to those of ordinary skill in the art that variations in the preferred devices and methods may be used and that it is intended that the invention may be practiced otherwise than as specifically described herein. Accordingly, this invention includes all modifications encompassed within the spirit and scope of the invention as defined by the claims that follow.

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  • Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Analytical Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Hematology (AREA)
  • Clinical Laboratory Science (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
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Abstract

Plaque comportant plusieurs cellules de réaction de taille uniforme réalisées sur la face supérieure, la densité de ces cellules de réaction étant d'au moins 10 par cm2. De préférence, la surface de chacune des ouvertures de ces cellules de réaction n'excède pas 55 % environ de la surface définie par le produit de multiplication de (1) l'espace compris entre les cellules de réaction de rangées différentes et (2) l'espace compris entre les cellules de réaction de colonnes différentes.
EP97917901A 1996-04-09 1997-04-09 Plaque pour systeme de reaction Withdrawn EP0892672A4 (fr)

Applications Claiming Priority (7)

Application Number Priority Date Filing Date Title
US630018 1996-04-09
US08/630,018 US5840256A (en) 1996-04-09 1996-04-09 Plate for reaction system
US73063696A 1996-10-11 1996-10-11
US730636 1996-10-11
US08/744,386 US6033544A (en) 1996-10-11 1996-11-07 Liquid distribution system
US744386 1996-11-07
PCT/US1997/005841 WO1997037755A1 (fr) 1996-04-09 1997-04-09 Plaque pour systeme de reaction

Publications (2)

Publication Number Publication Date
EP0892672A1 true EP0892672A1 (fr) 1999-01-27
EP0892672A4 EP0892672A4 (fr) 2001-02-28

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EP97917901A Withdrawn EP0892672A4 (fr) 1996-04-09 1997-04-09 Plaque pour systeme de reaction

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EP (1) EP0892672A4 (fr)
JP (1) JP2000508961A (fr)
AU (1) AU2609997A (fr)
CA (1) CA2251160A1 (fr)
WO (1) WO1997037755A1 (fr)

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EP1123499A2 (fr) * 1998-09-22 2001-08-16 Cellomics, Inc. Methodes et appareil de criblage sur la base de cellules avec un reseau miniaturise de cellules
US6942771B1 (en) 1999-04-21 2005-09-13 Clinical Micro Sensors, Inc. Microfluidic systems in the electrochemical detection of target analytes
US8481268B2 (en) 1999-05-21 2013-07-09 Illumina, Inc. Use of microfluidic systems in the detection of target analytes using microsphere arrays
US6908594B1 (en) 1999-10-22 2005-06-21 Aclara Biosciences, Inc. Efficient microfluidic sealing
JP2004500233A (ja) * 1999-10-22 2004-01-08 アクララ　バイオサイエンシーズ，　インコーポレイテッド 微小流体デバイスのためのシーリング
US6875619B2 (en) 1999-11-12 2005-04-05 Motorola, Inc. Microfluidic devices comprising biochannels
US6742661B1 (en) * 2001-04-03 2004-06-01 Micronics, Inc. Well-plate microfluidics
EP1636017A2 (fr) 2003-05-20 2006-03-22 Fluidigm Corporation Procede et systeme pour dispositif microfluidique et son imagerie

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WO1995027196A1 (fr) * 1994-04-04 1995-10-12 Sanadi Ashok R Procede et appareil destine a empecher la contamination croisee de plaques de test a cupules multiples
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JP2000508961A (ja) 2000-07-18
EP0892672A4 (fr) 2001-02-28
AU2609997A (en) 1997-10-29
CA2251160A1 (fr) 1997-10-16
WO1997037755A1 (fr) 1997-10-16

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