Apparatus and methods for efficient processing of biological samples on slides

• US 6,703,247 B1
• Filed: 12/23/1998
• Issued: 03/09/2004
• Est. Priority Date: 12/23/1996
• Status: Expired due to Fees

• Current Assignee: American Registry LLC
• Original Assignee: American Registry LLC
• Inventors: Chu, Wei-Sing
• Primary Examiner(s): Soderquist, Arlen
• Application Number:
  US09/219,443
• Time in Patent Office: 1,903 Days
• Field of Search: 422/99, 422/55, 422/100, 422/56, 422/102, 422/58, 422/104, 422/61, 422/68.1, 435/287.2, 435/287.9, 435/288.3, 435/288.4, 436/46, 436/180, 427/4, 427/2.11, 427/2.13
• US Class Current: 436/180
• 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: C12Q 1/6841 In situ hybridisation
• CPC Class Codes: C12Q 1/686 Polymerase chain reaction [PCR]
• CPC Class Codes: C12Q 2543/101 in situ amplification
• CPC Class Codes: G01N 1/312 for samples mounted on planar substrates
• CPC Class Codes: Y10T 436/112499 with sample on test slide
• CPC Class Codes: Y10T 436/2575 Volumetric liquid transfer

Abstract

Slideholders which are useful for manually or automatically processing biological samples on microscope slides are described. These slideholders hold multiple slides and are designed in conjunction with specialized trays for rapidly processing the mounted biological samples such as for immunocytochemical staining. The slideholder plus tray assemblies incorporate several useful advantages including a requirement for minimal reaction fluid volumes, ease of handling several slides concurrently, prevention of evaporation of reaction fluids, protection of the biological sample from extraneous environmental contamination, and the ability to perform in situ PCR. Various aspects of the design aid in removing trapped air from the reaction fluids and in adding fluids to the biological sample. One embodiment comprises a coverstip with a soft top which aids in prevention of tissue degradation by preventing pressure buildup during PCR. The system results in very low background signals and allows for manually processing manifold times the number of slides as is typically possible with other current manual methods. Another aspect of the invention is the use of predried reagents in wells, especially the use of predried reagents which dissolve sequentially. Yet another aspect of the invention is the use of external controls placed directly on a microscope slide in conjunction with a biological sample to be assayed. The external controls can be conveniently placed on a membrane which can be affixed to the slide. A further aspect of the invention is a specially designed tray to allow whole chromosome painting of all chromosomes of a cell sample on a single slide.

Claims

1. A method for treating a biological sample with a reagent or reagents, said method consisting essentially of:

(a) mounting said biological sample onto a microscope slide,

(b) inserting said microscope slide into a slideholder, said slideholder being capable of holding a plurality of slides,

(c) predrying at least one reagent in a well on a tray,

(d) placing said microscope slide in said slideholder sample side down on top of said well to cover said well, and

(e) filling said well completely with water or buffer to dissolve said reagent or reagents and allow said biological sample to contact said reagent or reagents in said well, and wherein capillary action of the dissolved reagent or reagents in said well contacting the microscope slide effectively seals the well from the atmosphere.

2. The method of claim 1, wherein more than one reagent is predried in said well.

3. The method of claim 2, wherein reagents are dried sequentially in reverse order of action.

4. The method of claim 3, wherein said reagents are separated from each other by an inert material.

5. A method for treating a biological sample with a reagent or reagents, said method consisting essentially of:

(a) mounting said biological sample onto a microscope slide,

(b) inserting said microscope slide into a slideholder, said slideholder being capable of holding a plurality of slides,

(c) filling a well on a tray completely with a solution of a reagent or reagents, and

(d) placing said microscope slide in said slideholder sample side down on top of said well to cover said well, allowing said biological sample to contact said solution of reagent or reagents in said well, and wherein capillary action of said solution of a reagent or reagents in said well contacting the microscope slide effectively seals the well from the atmosphere.

6. The method of claim 5, wherein said solution of a reagent or reagents contains an immunocytochemical staining reagent or reagents.

7. The method of claim 5, wherein said solution of a reagent or reagents contains a histochemical staining reagent or reagents.

8. The method of claim 5, wherein said solution of a reagent or reagents contains a reagent or reagents for in situ hybridization.

9. The method of claim 5, wherein said solution of a reagent or reagents contains a reagent or reagents for in situ polymerase chain reaction.

10. A method for treating a biological sample with a reagent or reagents, said method consisting essentially of:

(a) mounting said biological sample onto a microscope slide,

(b) filling a well on a tray completely with a solution of a reagent or reagents, and

(c) placing said microscope slide sample side down on top of said well to cover said well, allowing said biological sample to contact said solution of reagent or reagents in said well, and wherein capillary action of said solution of a reagent or reagents in said well contacting the microscope slide effectively seals the well from the atmosphere.

11. The method of claim 10, wherein said solution of a reagent or reagents contains an immunocytochemical staining reagent or reagents.

12. The method of claim 10, wherein said solution of a reagent or reagents contains a histochemical staining reagent or reagents.

13. The method of claim 10, wherein said solution of a reagent or reagents contains a reagent or reagents for in situ hybridization.

14. The method of claim 10, wherein said solution of a reagent or reagents contains a reagent or reagents for in situ polymerase chain reaction.