Manifold vacuum device for biochemical and immunological uses

• US 4,777,021 A
• Filed: 03/19/1987
• Issued: 10/11/1988
• Est. Priority Date: 04/25/1986
• Status: Expired due to Fees

• Current Assignee: Wertz Richard K
• Original Assignee: Richard K. Wertz
• Inventors: Watkins, Linda R., Wertz, Richard K.
• Primary Examiner(s): Scott, Samuel
• Assistant Examiner(s): KAMEN, NOAH P
• Application Number:
  US07/027,827
• Time in Patent Office: 572 Days
• Field of Search: 435/284, 435/285, 435/293, 435/300, 435/301, 435/311, 435/299, 422/101, 422/100, 436/177, 436/809, 436/178, 210/406, 210/323.1, 210/500.28
• US Class Current: 422/534
• CPC Class Codes: B01L 3/50255 Multi-well filtration
• CPC Class Codes: C12M 23/12 Well or multiwell plates C12M25/04 takes precedence
• CPC Class Codes: C12M 23/40 Manifolds; Distribution pieces fluid transfer means B01L3/563
• CPC Class Codes: Y10T 436/255 including use of a solid sorbent, semipermeable membrane, or liquid extraction

Abstract

A device for simultaneously performing multiple biochemical or immunological reactions. A manifold plate rests atop a waste fluid collection chamber or receptacle suitable for attachment to a vacuum pump. The manifold contains a series of wells that serve as reaction vessels. The sides of each well are fluid-impermeable, and its lower end provides a planar, annular base. A hydrophobic membrane is sealed to the lower surface of the base of each well. The membrane is formed of a material through which fluid can be drawn upon application of vacuum while retaining the fluid within the well in the absence of vacuum. By placement of a solid-phase support for binding biological coreactants atop the membrane prior to initiation of the reaction, solid-phase reaction products can be collected when vacuum is applied and can then be freely withdrawn from its well. The membranes of all of the reaction wells are physically isolated from each other so that totally independent biochemical or immunological reactions can be performed in each well without danger of cross-contamination between wells.

Claims

1. A microtiter tray for use with a vacuum chamber therebelow, comprising:

a manifold plate formed to define a series of substantially cylindrical, wells with fluid-impermeable walls having inner and outer surfaces, each said wall having a planar, annular base at its lower end surrounding a circular bottom opening of said well the full diameter across said wall'"'"'s upper surfaces,

a series of separate hydrophobic, liquid-permeable membranes means, each secured to and sealed to the lower surface of a said base of one said well, for closing the lower end of said well along a flat plane and for enabling said wells to contain a reagent sample to be tested, without leakage so long as no significant pressure differential exists between the upper and lower surfaces of said membrane means and said membrane means'"'"' lower surface is retained free from contact with an absorbent material, said membrane means affording passage of said sample under significant pressure differential and when in contact with absorbent material, each said membrane means having an outer periphery separated from the outer periphery of all adjacent wells, so that fluid cannot migrate from one to another, and having a solid-phase means for supporting a biological coreactant for bonding thereto a reaction product resulting from said reagent, contained in said well above and itself supported by said membrane means.

2. The microtiter tray of claim 1 wherein the outer periphery of each said membrane means extends out no further than the outer surface of the walls of its well.

3. The microtiter tray of claim 1 wherein said membrane means is a spun-bonded polyester in a sheet of compacted continuous-filament polyester fibers.

4. A inexpensive, disposable microtiter tray for use with a vacuum chamber therebelow, comprising:

a manifold plate formed of plastic to define a series of fluid-impermeable substantially cylindrical wells fully open at their lower ends, each well having a cylindrical well with a planar, annular base at its lower end, and

a series of separate hydrophobic, liquid-permeable membranes, each permanently secured to and sealed to the lower surface of a said planar base of one said well to close said well for temporary containment of a liquid comprising a reagent sample nonreactive with said plastic and said membrane during incubation and the like, while being sufficiently spaced from the membrane of all adjacent well to prevent fluid migration thereto, and

each said membrane providing passage through it of said liquid without binding a reagent from said liquid, when subjected to significant pressure differential above and below said membrane, and a solid-phase porous fiber means within atleast some of said wells, and freely withdrawable therefrom, each above and detached from a said membrane, for supporting a biological coreactant for bonding to a reaction product of said reagent with said correaction.

5. The microtiter tray of claim 4 in which each said membrane has an outer periphery substantially coincident with that of its well.

6. The microtiter tray of claim 4 wherein said membrane is a spun-bonded polyester in a sheet of compacted continuous-filament polyester fibers.

7. A microtiter tray-vacuum chamber combination comprising:

a manifold plate formed to define a series of fluid-impermeable wells having substantially cylindrical walls and fully open at their lower ends, each wall having a planar, annular base portion at its lower end, and

a series of separate hydrophobic, liquid-permeable membrane means each secured to and sealed to the lower surface of a said base of one said well, for closing the lower end of said well along a flat plane and for enabling said wells to contain a reagent sample to be tested without leakage so long and only so long as no significant pressure differential exists between the upper and lower surfaces of said membrane and said membrane'"'"'s lower surface is retained free from contact with an absorbent material, each said membrane means having an outer periphery separated from the outer periphery of all adjacent wells, so that fluid cannot migrate from one to another,

a lower receptacle beneath said manifold plate providing a vacuum chamber with an imperforate floor underlying said plate, said lower receptacle providing an outlet from said vacuum chamber, and

a gasket sealing said plate to said lower receptacle and enclosing the vacuum chamber.

8. The combination of claim 7, wherein said lower receptacle provides a floor for said vacuum chamber that slopes toward said outlet.

9. The combination of claim 7 having solid-phase porous, fiber means within, and freely withdrawable from, at least some of said wells, each above and detached from a said membrane, for supporting a biological coreactant for bonding to a reaction product of said reagent with said coreaction.

10. The combination of claim 7 wherein said membrane is a spun-bonded polyester in a sheet of compacted continuous-filament polyester fibers.

11. A laboratory assembly, comprising:

a manifold plate formed of plastic to define a series of small volume, fluid-impermeable, smooth-bore wells, each well having a capacity of about 25-100 microliters and being fully opened at its lower end and providing a planar, annular base at its lower end,

a series of separate hydrophobic, liquid-permeable membranes, each permanently secured to and sealed to the lower surface of a said planar base of one said well to close said well for temporary containment of a liquid comprising reagent samples nonreactive with said plastic and said membrane during incubation and the like, while being sufficiently spaced from the membranes of all adjacent wells to prevent liquid migration thereto,

each said membrane providing passage through it of said liquid without binding a reagent from said liquid, when subjected to significant pressure differential above and below said membrane, and

solid-phase support means for bonding biological coreactants, supported loosely in each said well by a said membrane for binding a selected reagent from a said liquid inserted in its said well,

a lower receptacle beneath said manifold plate providing a vacuum chamber underlying said plate, said receptacle providing an outer from said vacuum chamber, said receptacle providing an imperforate floor for said vacuum chamber,

a gasket sealing said plate to said receptacle and enclosing the vacuum chamber,

a source of vacuum suction of sufficient strength to draw the liquid of said reagent samples through the fluid-permeable membranes leaving a reactant bonded to said coreactants on said solid-phase support means in said wells, and

connecting means leading from said vacuum chamber to said vacuum source.

12. The device of claim 11 in which said floor slopes toward said outlet.