Apparatus and methods for processing biological samples and a reservoir therefor

• US 8,632,739 B2
• Filed: 05/16/2012
• Issued: 01/21/2014
• Est. Priority Date: 01/08/2004
• Status: Active Grant

• Current Assignee: Agilent Technologies, Inc.
• Original Assignee: Dakocytomation California Incorporated
• Inventors: Testa, Gregory A., Poulsen, Tim Svenstrup, Matthiesen, Steen Hauge, Rasmussen, Ole Feldballe, Winther, Lars
• Primary Examiner(s): Jarrett, Lore
• Application Number:
  US13/472,777
• Publication Number:
  US 20130023038A1
• Time in Patent Office: 615 Days
• Field of Search: 422/547
• US Class Current: 422/547
• CPC Class Codes: B01L 2200/142 Preventing evaporation
• CPC Class Codes: B01L 2300/043 Hinged closures
• CPC Class Codes: B01L 2300/069 Absorbents; Gels to retain a fluid
• CPC Class Codes: B01L 2300/0822 Slides
• CPC Class Codes: B01L 2300/10 Means to control humidity and/or other gases
• CPC Class Codes: B01L 2300/1822 using Peltier elements
• CPC Class Codes: B01L 2300/1844 using fans
• CPC Class Codes: B01L 3/508 rigid containers not provided for above
• CPC Class Codes: B01L 7/52 with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples amplification or hybridisation processes per se C12Q1/68; controlling sequential reactions for synthesis B01J19/0046
• CPC Class Codes: C12M 41/14 Incubators; Climatic chambers per se B01L1/00
• CPC Class Codes: C12M 41/36 of biomass, e.g. colony counters or by turbidity measurements electrooptical investigation of individual particles G01N15/14, flow cytometers G01N15/1404
• CPC Class Codes: C12Q 1/6841 In situ hybridisation
• CPC Class Codes: G01N 1/312 for samples mounted on planar substrates
• CPC Class Codes: Y10T 436/143333 Saccharide [e.g., DNA, etc.]

Abstract

An apparatus for processing at least one biological sample accommodated on at least one carrier member (15) in a chamber includes, at least one reservoir (18) able to accommodate a fluid on a surface inside the chamber adjacent to and/or facing a substantial part of the at least one biological sample. The apparatus may comprise a bottom member (12) arranged to support at least one carrier member (15) carrying at least one biological sample and a lid (14) including at least one fluid reservoir (18). The reservoir filled with water provides humidity to the chamber and impedes drying out of the sample.

Claims

1. An apparatus adapted for nucleic acid denaturization and hybridization of at least one biological sample on at least one carrier member in a chamber, said apparatus comprising:

a bottom member supporting said at least one carrier member;

a hydrophilic porous solid reservoir configured to sorb and desorb water and water vapor;

a lid for the chamber, detachable from the bottom member, the lid and the bottom member forming a closed chamber when the lid is positioned atop the bottom member, wherein the lid is configured to position the reservoir inside the closed chamber directly above the at least one biological sample such that the reservoir does not touch said at least one biological sample, when said lid is positioned atop the bottom member to form the closed chamber; and

a sample temperature controlling device below said at least one carrier member controlling the temperature of said at least one carrier member and said at least one biological sample, wherein said reservoir either sorbs or desorbs water and water vapor so as to control relative humidity within said chamber at above 85% relative humidity, when said lid is positioned atop the bottom member to form the closed chamber.

2. The apparatus of claim 1, wherein said lid may be removed from the bottom member to access said at least one biological sample and said reservoir.

3. The apparatus of claim 1, wherein said reservoir is less than 5 cm from said at least one carrier member.

4. The apparatus of claim 1, wherein said reservoir is less than 1.0 cm from said at least one carrier member.

5. The apparatus of claim 1, wherein said reservoir is less than 0.5 cm from said at least one carrier member.

6. The apparatus of claim 1, wherein a macroscopic surface area of said reservoir is greater than 10% of the total area of said at least one carrier member.

7. The apparatus of claim 1, wherein said sample temperature controlling device further comprises at least one element selected from the group consisting of a temperature sensor, heating wires, an inductive heater, and a cooling device for cooling the at least one biological sample.

8. The apparatus of claim 1, wherein relative humidity in the closed chamber is controlled by desorption of water and water vapor from said reservoir during heating of said at least one biological sample or absorption of water and water vapor from said closed chamber during cooling of said at least one biological sample.

9. The apparatus of claim 1, wherein said reservoir is shaped as a substantially flat sheet.

10. The apparatus of claim 9, wherein the thickness of said reservoir is less than 10% of the length of said reservoir.

11. The apparatus of claim 1, wherein said water further comprises at least one additive, the at least one additive selected from the group consisting of an anti-microbial agent, an inorganic salt, a detergent and an organic solvent.

12. The apparatus of claim 1, wherein said reservoir is made of a material from the group consisting of polymeric fiber composites, polymeric fiber blends, glass fiber materials, expanded porous polymers, porous ceramics, wood pulp, cardboard, leather and cellulose-based materials, or is made of a material comprising a composition chosen from the group consisting of polyethylene, polypropylene, polyurethanes, polysulfones, polyvinyl, polyacrylics, ethylene Vinyl Acetate, viscose rayon, polystyrene, macroreticular polystyrene, aliphatic polymers, phenol-formaldehyde condensate polymers, epoxy, cotton, polysaccharide, modified polysaccharides, wood pulp, calcium carbonate, silica gels, glass fiber, bentonite, perlite and zeolite.

13. The apparatus of claim 1, wherein said reservoir is made of a material comprising bundles of fibers or loose material chosen from the group consisting of polymeric bonded fibers, non-bonded fibers, woven fibers, knitted fibers, micro fibers, textiles, tufted textiles, bonded polyamide fibers, polyester fibers, polyolefin fibers, cellulose acetate fibers, and non-woven and bonded blends of hydrophilic modified polypropylene and polyethylene micro fibers, wherein the bundles of fibers or loose material are retained by a thin wall of film.

14. The apparatus of claim 1, wherein said reservoir is impregnated with an anti microbial agent or other protective agent.

15. The apparatus of claim 1, wherein the type, shape and size of material of which said reservoir is comprised is selected to optimize surface properties of said material with regard to a surface tension of said water.

16. The apparatus of claim 1, wherein material of which said reservoir is comprised has a density from 0.050 to 1.5 gram/cm³.

17. The apparatus claim 1, wherein said reservoir has the ability to hold at least a predefined minimum volume of liquid per carrier member, the predefined minimum volume being chosen from the group consisting of 10 micro-liters, 100 micro-liters, 200 micro-liters, 500 micro-liters, and 1000 micro-liters.

18. The apparatus of claim 1, further comprising a second reservoir outside the closed chamber.

19. A method of processing biological samples, wherein at least one biological sample is arranged on a carrier member, for treatment to prepare the sample by staining, the method comprising the steps of:

providing a bottom member for supporting said carrier member;

positioning a lid, detachable from the bottom member, above the carrier member to form a closed chamber with the bottom member, wherein positioning the lid above the carrier member also positions a hydrophilic porous solid reservoir configured to sorb and desorb water and water vapor directly above the at least one biological sample such that the reservoir does not touch said at least one biological sample;

controlling the temperature of said carrier member and said at least one biological sample with a sample temperature controlling device positioned below said carrier member; and

maintaining at least 85% relative humidity inside the chamber through the sorption or desorption of water and water vapor.