Method for processing non-liquid biological samples with dynamic application of a processing liquid

• US 8,501,434 B2
• Filed: 05/03/2012
• Issued: 08/06/2013
• Est. Priority Date: 10/06/2010
• Status: Active Grant

• Current Assignee: Biocare Medical LLC
• Original Assignee: Biocare Medical LLC
• Inventors: Barker, Stephen, Avantsa, Saradha
• Primary Examiner(s): Gakh, Yelena G
• Application Number:
  US13/463,500
• Publication Number:
  US 20120214191A1
• Time in Patent Office: 460 Days
• Field of Search: None
• US Class Current: 435/40.5
• CPC Class Codes: B01F 31/65 the materials to be mixed being directly submitted to a pulsating movement, e.g. by means of an oscillating piston or air column
• CPC Class Codes: B01F 33/3035 using surface tension to mix, move or hold the fluids
• CPC Class Codes: G01N 1/31 Apparatus therefor
• CPC Class Codes: G01N 1/312 for samples mounted on planar substrates
• CPC Class Codes: G01N 33/54346 Nanoparticles
• CPC Class Codes: G01N 33/587 Nanoparticles
• CPC Class Codes: G01N 35/0098 involving analyte bound to insoluble magnetic carrier, e.g. using magnetic separation magnetic particles used in immunoassays G01N33/54326; magnetic separation in general B03C
• CPC Class Codes: Y10T 436/25 including sample preparation

Abstract

Systems and methods of sample (1) and staining processing including compression and dynamic movement of liquids (3) in a fluidically moving substantially contained liquid bridge (6) perhaps between a hydrophobic wand (4) and a hydrophilic sample support element (2). Embodiments may include low volume reagent and perhaps even low volume buffer wash in sample processing. In addition, antibodies can be conjugated with nanoparticles (64) and can be used in sample processing. Exposing a sample with or without movement to AC, DC, or even a permanent magnet field may improve staining. Staining with nanoparticle reagents could be quantified using a microscope with a magnetometer below the slide viewing area. The detection of nanoparticles attached to the chemistry may facilitate the quantification of cancerous cells stained in the tissue.

Claims

1. A method for efficient processing of a non-liquid biological sample with a dynamically moving aqueous-based processing liquid, comprising the steps of:

providing a stationary sample support with a hydrophilic surface;

providing the non-liquid biological sample fixed on the stationary sample support;

providing a moveable wand positioned above the sample support element, the movable wand having a substantially flat hydrophobic surface;

applying the aqueous-based processing liquid to the stationary sample support with the non-liquid biological sample;

contacting said substantially flat hydrophobic surface of the moveable wand with the aqueous-based processing liquid;

forming a substantially contained liquid bridge with the aqueous-based processing liquid between the substantially flat hydrophobic surface of the wand and the hydrophilic surface of the stationary sample support;

steadily oscillating the moveable wand back and forth above the stationary sample support and the non-liquid biological sample, therefore steadily oscillating the substantially contained liquid bridge over the stationary sample support and the non-liquid biological sample; and

efficiently processing the non-liquid biological sample with the aqueous-based processing liquid.

2. The method according to claim 1 wherein the step of steadily oscillating the moveable wand back and forth is performed with a constant velocity.

3. The method according to claim 1 wherein the step of steadily oscillating the moveable wand back and forth is performed with a variably velocity.

4. The method according to claim 1 wherein the substantially contained liquid bridge comprises a meniscus at each of its sides.

5. The method according to claim 1 further comprising compressing the oscillating liquid bridge between the wand and the sample support thus forming sliding reagent friction between the oscillating liquid bridge and the non-liquid biological sample.

6. The method according to claim 1 wherein the non-liquid biological sample is selected from a group consisting of a untreated tissue, pretreated tissue, antigen retrieved tissue, epitope retrieved tissue, deparaffinized tissue, histological sample, cell specimens, cell lines, cell membrane, cell preparations, proteins, synthetic peptides, bodily fluids, bone marrow, cytology specimens, blood smears, and any combination thereof.

7. The method according to claim 1 wherein the stationary sample support comprises a glass slide.

8. The method according to claim 1 wherein the step of steadily oscillating the moveable wand back and forth comprises providing that each movement of the substantially contained liquid bridge covers all of the non-liquid biological sample.

9. The method according to claim 1 wherein the step of efficiently processing the non-liquid biological sample comprises the step of staining said sample.

10. The method according to claim 9 wherein the step of staining the non-liquid biological sample provides a stained sample having a property selected from a group consisting of a sharp stain, crisp stain, a stained sample with substantially no background, a stained sample with substantially no non-specific staining, and a stained sample with substantially no hue on the sample support element.

11. The method according to claim 1, wherein forming a substantially contained liquid bridge with the aqueous-based processing liquid between the substantially flat hydrophobic surface of the wand and the hydrophilic surface of the stationary sample support comprises forming an acute angle between the liquid bridge and the hydrophilic surface of the stationary sample support and an obtuse angle between the liquid bridge and the hydrophobic surface of the wand.

12. The method according to claim 1 wherein the substantially flat hydrophobic surface comprises a rough hydrophobic surface.

13. The method according to claim 1 wherein the aqueous-based processing liquid comprises a reagent.

14. The method according to claim 13 wherein the reagent is selected from a group consisting of an unstable reagent, a stable reagent, a immunohistochemistry reagent, an in situ hybridization reagent, a histochemical reagent, a pretreatment reagent, a reagent for deparaffinization, a reagent for antigen retrieval, and any combination thereof.

15. The method according to claim 13 wherein the reagent comprises at least one component selected from a group consisting of antibodies, probes, antibody-enzyme conjugates, enzymes, nanoparticles, and any combination thereof.

16. The method according to claim 1 wherein the aqueous-based processing liquid comprises a buffer.

17. The method according to claim 16 wherein the buffer comprises a wash buffer.

18. The method according to claim 1 wherein the aqueous-based processing liquid comprises a stain.

19. The method according to claim 18 wherein the stain comprises at least one component selected from a group consisting of chromogenic substrates, chromogenic activator, chromogens, counterstains, and any combination thereof.