Removal of embedding media from biological samples and cell conditioning on automated staining instruments

• US 6,544,798 B1
• Filed: 05/11/2001
• Issued: 04/08/2003
• Est. Priority Date: 02/26/1999
• Status: Expired due to Term

• Current Assignee: Ventana Medical Systems Incorporated (Roche Holding AG)
• Original Assignee: Ventana Medical Systems Incorporated (Roche Holding AG)
• Inventors: Ward, Glen, Christensen, Kimberly, Sebastiao, Noemi, Macrea, Ethel R.
• Primary Examiner(s): WALLENHORST, MAUREEN
• Application Number:
  US09/853,200
• Time in Patent Office: 697 Days
• Field of Search: 436/8, 436/17, 436/18, 436/63, 436/175, 436/177, 435/40.5, 435/40.51, 435/40.52, 435/962
• US Class Current: 436/177
• CPC Class Codes: G01N 1/36 Embedding or analogous mounting of samples
• CPC Class Codes: Y10T 436/107497 Preparation composition [e.g., lysing or precipitation, etc.]
• CPC Class Codes: Y10T 436/25125 Digestion or removing interfering materials
• CPC Class Codes: Y10T 436/25375 Liberation or purification of sample or separation of material from a sample [e.g., filtering, centrifuging, etc.]

Abstract

The present invention provides reagents for use in an automated environment for cell conditioning of biological samples wherein the cells or tissues are predisposed for access by reagent molecules for histochemical and cytochemical staining procedures. The reagents include components optimized to facilitate molecular access to cells and cell constituents within the biological sample. The present invention also provides reagents for use in an automated environment for removing or etching embedding media by exposing a biological sample to be stained in histochemical or cytochemical procedures without the dependence on organic solvents. The reagents include components optimized to facilitate removal or etching of the embedding media from the biological sample.

Claims

1. An automated method of removing embedding media from a biological sample on a microscope slide, the method comprising the sequential steps of:

a) separately heating a fluid, the fluid being immiscible with the embedding media; and

b) contacting said biological sample with said heated immiscible fluid thereby heating the embedding media sufficient to separate the embedding media from the biological sample.

2. The method of claim 1, wherein the step of heating the immiscible fluid heats the immiscible fluid to a temperature above a melting point of the embedding media.

3. The method of claim 2, wherein said heating step requires the temperature of said immiscible fluid to be at least about 65 degrees centigrade.

4. The method of claim 3, wherein said embedding media is paraffin or paraffin-based.

5. The method of claim 2, wherein contacting the biological sample is carried out by dispensing said immiscible fluid onto the biological sample and maintaining the temperature of the immiscible fluid above the melting point of the embedding media.

6. The method of claim 5, wherein maintaining the temperature is effected by recirculating said immiscible fluid.

7. The method of claim 2, further comprising the step of heating the biological sample and the embedding media.

8. The method of claim 7, wherein the step of heating the biological sample and the embedding media includes heating the biological sample and the embedding media above a melting point of the embedding media.

9. The method of claim 1, wherein contacting the biological sample includes rinsing the biological sample with said heated immiscible fluid.

10. The method of claim 9, wherein the step of rinsing the biological sample with said heated immiscible fluid comprises rising the biological sample with a continuous stream of the heated immiscible fluid.

11. The method of claim 1, further comprising the step of heating the biological sample and the embedding media.

12. The method of claim 11, wherein the step of heating the biological sample and the embedding media includes heating the biological sample and the embedding media above a melting point of the embedding media.

13. The method of claim 1, wherein separating the embedding media from the biological sample includes causing the embedding media to float on the immiscible fluid so as to be removed.

14. The method of claim 1, wherein the immiscible fluid comprises water.

15. The method of claim 1, wherein the immiscible fluid is selected from the group consisting of de-ionized water, citrate buffer (pH 6.0-8.0), Tris-HCl buffer (pH 6-10), phosphate buffer (pH 6.0-8.0), SSC buffer, acidic buffers or solutions (pH 1-6.9), basic buffers or solutions (pH 7.1-14), mineral oil, Norpar, canola oil, and PAG oil.

16. The method of claim 1, wherein the immiscible fluid contains ionic or non-ionic surfactants.