Miniaturized biological assembly

• US 5,200,152 A
• Filed: 12/27/1990
• Issued: 04/06/1993
• Est. Priority Date: 03/28/1988
• Status: Expired due to Term

• Current Assignee: Cytonix LLC (Thermo Fisher Scientific Incorporated)
• Original Assignee: Cytonix Corporation (Thermo Fisher Scientific Incorporated)
• Inventors: Brown, James F.
• Primary Examiner(s): KRYZA, LYNN M
• Application Number:
  US07/632,655
• Time in Patent Office: 831 Days
• Field of Search: 422/58, 422/68.1, 422/100, 422/102, 422/61, 422/101, 436/46, 436/165, 436/809, 356/244, 356/246, 435/285, 435/300, 435/301, 435/310, 435/810, 435/970
• US Class Current: 422/503
• CPC Class Codes: B01L 2200/0689 Sealing
• CPC Class Codes: B01L 2200/142 Preventing evaporation
• CPC Class Codes: B01L 2300/0822 Slides
• CPC Class Codes: B01L 2300/0887 Laminated structure
• CPC Class Codes: B01L 2400/0406 capillary forces
• CPC Class Codes: B01L 3/5027 by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip B01L3/5023 takes precedence; micromixers B01F33/30; microreactors for synthesis B01J19/0093; microcapillary devices in general B81B1/00
• CPC Class Codes: B01L 3/502707 characterised by the manufacture of the container or its components by shaping or joining plastic parts B29C59/00, B29C65/00; by laminating B32B37/00; manufacture of microstructural devices in general B81C
• CPC Class Codes: Y10S 435/81 Packaged device or kit
• CPC Class Codes: Y10S 436/809 Multifield plates or multicontainer arrays
• CPC Class Codes: Y10T 436/112499 with sample on test slide

Abstract

A miniature chamber assembly that provides a miniature capillary environment in which a liquid medium containing microscopic-size particulate material can be placed for study under a microscope. The assembly includes components which do not wet relative to the liquid medium. The assembly provides a miniaturized capillary environment that can contain the liquid medium for a period of time sufficient to enable observation while preventing deterioration of the medium.

Claims

1. A miniaturized biological assembly for containing a sample of microscopic-sized particulate biological material in a fluid medium to enable quantitative microscopic examination thereof, said assembly comprising

first and second plates which are disposed in registry with one another and adhered to one another,

at least one patterned layer located between said first and second plates to define at least one chamber between said first and second plates for a sample of particulate biological material in a fluid medium and to define a boundary which minimizes contamination and drying of liquid sample and protects liquid sample against deterioration at temperatures at which liquid sample is maintained for study, said at least one chamber having a depth dimension between said first and second plates which enables all particulate biological material therein to be viewed within a particular depth of field characteristic of a microscope,

attachment means to fixedly position said first and second plates together with said at least one patterned layer therebetween, and

a plurality of annular pads located within said at least one chamber and extending between said first and second plates, and wherein said attachment means comprises adhesive within said annular pads to adhere said first and second plates together.

2. A miniaturized biological assembly for containing a sample of microscopic-sized particulate biological material in a fluid medium to enable quantitative microscopic examination thereof, said assembly comprising

first and second plates which are disposed in registry with one another and adhered to one another,

at least one patterned layer located between said first and second plates to define at least one chamber between said first and second plates for a sample of particulate biological material in a fluid medium and to define a boundary which minimizes contamination and drying of liquid sample and protects liquid sample against deterioration at temperatures at which liquid sample is maintained for study, said at least one chamber having a depth dimension between said first and second plates which enables all particulate biological material therein to be viewed within a particular depth of field characteristic of a microscope, and

attachment means to fixedly position said first and second plates together with said at least one patterned layer therebetween,

wherein said at least one patterned layer defines a first end boundary having a notch, opposite side boundaries, a second end boundary opposite said first end boundary, and two parallel legs which extend from said second end boundary into said notch so as to define a sample channel between said two legs and first and second sample chambers on opposite sides of said legs.

3. A miniaturized biological assembly as defined in claim 2, wherein said second plate has a smaller length than said first plate so that said notch is exposed to an environment around said miniaturized assembly and provides an aperture for supply of sample to said sample channel and said first and second sample chambers.

4. A miniaturized biological assembly as defined in claim 3, wherein said sample channel has a width of about 2 mm and said legs define entrance passages to said first and second sample chambers having widths of about 2 mm.

5. A miniaturized biological assembly for containing a sample of microscopic-sized particulate biological material in a fluid medium to enable quantitative microscopic examination thereof, said assembly comprising

first and second plates which are disposed in registry with one another and adhered to one another,

at least one patterned layer located between said first and second plates to define at least one chamber between said first and second plates for a sample of particulate biological material in a fluid medium and to define a boundary which minimizes contamination and drying of liquid sample and protects liquid sample against deterioration at temperatures at which liquid sample is maintained for study, said at least one chamber having a depth dimension between said first and second plates which enable all particulate biological material therein to be viewed within a particular depth of field characteristic of a microscope, and

attachment means to fixedly position said first and second plates together with said at least one patterned layer therebetween,

wherein said at least one patterned layer is a patterned resist.

6. A miniaturized assembly for containing a sample of liquid or microscopic-sized particulate biological or chemical material in a fluid medium to enable microscopic examination thereof, said assembly comprising:

first and second plates which are disposed in registry with one another, at least one of said plates being sufficiently thin so as to allow proper focus by a microscope having a particular depth of focus over a depth of field thereof beyond an opposite side of said at least one of said plates, said first and second plates having a precise and sufficient thickness and modulus to provide a specific maximum deflection due to capillary forces created by the presence of a sample between said plates, the surfaces of said plates having proper electrostatic surface charge to limit adhesion of a sample, and said plates being free of particulate and chemical contamination;

at least one hydrophobic-oleophobic layer located in registry between said first and second plates to define at least one chamber for a sample, said at least one chamber formed by said at least one hydrophobic-oleophobic layer and opposed surfaces of said first and second plates, said at least one hydrophobic-oleophobic layer having a prescribed and accurate depth dimension, said depth dimension enabling all particulate material therein to be viewed within a depth of field characteristic of a microscope, said at least one hydrophobic-oleophobic layer also defining an aperture leading to said at least one chamber from the edge of at least one of said plates for the introduction of a sample into said at least one chamber, said at least one hydrophobic-oleophobic layer further defining at lest one channel for the venting of air displaced from said at least one chamber as it is filled with a sample, said at least one hydrophobic-oleophobic layer surrounding said at least one chamber and at least one channel to permit the escape of air trapped in said at least one chamber while preventing the flow of a sample through said at least one channel.

7. A miniaturized assembly as defined in claim 6, wherein at least one of said first and second plates is transparent to ultraviolet and/or visible light.

8. A miniaturized assembly as defined in claim 6, wherein at least one of said first and second plates is optically flat to less than 1 micrometer per cm.

9. A miniaturized assembly as defined in claim 6, wherein at least one of said first and second plates is angled at an edge opposing the surface of the other plate to form said aperture.

10. A miniaturized assembly as defined in claim 6, wherein attachment means are provided between the interior surfaces of the first and second plates.

11. A miniaturized assembly as define din claim 6, wherein a thin coating of hydrophobic material is adhered to at least portions of the exterior-facing surface of said first plate adjacent to said introduction aperture.

12. A miniaturized assembly as defined in claim 6, further including an attachment means to fixedly position said first and second plates together with said at least one hydrophobic-oleophobic layer therebetween, said attachment means providing a vapor seal at the perimeter of said at least one chamber except at said entrance aperture and said at least one vent channel, said attachment means restricting the size of said at least one vent channel to control the out-flow of air displaced from said at least one sample chamber by the introduction of a sample.

13. A miniaturized assembly as defined in claim 12, wherein said attachment means is disposed in registry with said first and second plates and said at least one hydrophobic-oleophobic layer to fixedly position said first and second plates together with said at least one hydrophobic-oleophobic layer therebetween, said attachment means being at least one member selected from the group consisting of screen-printed, ink-jet-printed attachment means and uv curable, pressure sensitive, solvent-free and melt-bonding adhesives.

14. A miniaturized assembly as defined in claim 6, further including a patterned transparent hydrophobic thin coating adhered to said at least one chamber surface of said first and second plates having properties that further limit the adhesion of a sample.

15. A miniaturized assembly as defined in claim 6, including at least one of said first and second plates smoothed to a radius of 5 to 50 micrometers at the edge opposing the surface of the other plate to limit mechanical damage to a sample during introduction through said aperture.

16. A miniaturized assembly as defined in claim 6, wherein said at least one hydrophobic-oleophobic layer has a surface energy and surface structure to produce an advancing contact angle of at least 140 degrees against water and air.

17. A miniaturized assembly as defined in claim 6, including a patterned layer having a specific predetermined thickness and having within-device, device-to-device, and lot-to-lot variation of less than ±

5% of a prescribed thickness.