Method for non-destructive macromolecule extraction from biological samples on slide

• US 20090286305A1
• Filed: 07/30/2009
• Published: 11/19/2009
• Est. Priority Date: 04/15/2005
• Status: Abandoned Application

• Current Assignee: Wei-Sing Chu
• Original Assignee: Wei-Sing Chu
• Inventors: Chu, Wei-Sing
• Application Number:
  US12/512,214
• Publication Number:
  US 20090286305A1
• Time in Patent Office: Days
• US Class Current: 435/270
• CPC Class Codes: B01D 11/02 of solids
• CPC Class Codes: B01D 11/0496 by extraction in microfluidic devices
• CPC Class Codes: B01L 2300/048 enabling gas exchange, e.g. vents
• CPC Class Codes: B01L 2300/0822 Slides
• CPC Class Codes: B01L 2300/0877 Flow chambers
• CPC Class Codes: B01L 2300/10 Means to control humidity and/or other gases
• CPC Class Codes: B01L 2300/1805 Conductive heating, heat from thermostatted solids is conducted to receptacles, e.g. heating plates, blocks
• CPC Class Codes: B01L 2400/0406 capillary forces
• CPC Class Codes: B01L 3/502753 characterised by bulk separation arrangements on lab-on-a-chip devices, e.g. for filtration or centrifugation separation in general B01D; microapparatus for analysis using electrophoresis G01N27/44791; sample preparation G01N1/28
• CPC Class Codes: G01N 1/28 Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Qmounting specimens on microscopic slides G02B21/34; means for supporting the objects or the materials to be analysed in electron microscopes H01J37/20 ; laboratory gas handling apparatus B01L5/00
• CPC Class Codes: G01N 1/312 for samples mounted on planar substrates
• CPC Class Codes: G01N 2001/4016 being a selective membrane, e.g. dialysis or osmosis
• CPC Class Codes: G01N 33/6803 General methods of protein analysis not limited to specific proteins or families of proteins

Abstract

A thin layer of a biological sample, such as a section of frozen or preserved tissue sample, a section of fresh or preserved cells, and a mono layer of prokaryotic and eukaryotic cells, is placed on a flat surface of a solid supporting base. Macromolecules, such as DNA, RNA, and proteins, are extracted directly from the thin layer of the biological sample that is attached to the supporting base.

Claims

1. A method for extracting biological molecules from a biological sample, comprising:

a) placing a thin layer of said biological sample onto a flat surface of a supporting base;

b) contacting said thin layer of said biological sample with a small volume of extraction solution, said extraction solution facilitates dissolution of predetermined biological molecules from said biological sample;

c) incubating at a temperature for a length of time; and

,

d) collecting said extraction solution which contains said predetermined biological molecules extracted from said biological sample.

2. A method according to claim 1, wherein incubation is performed in a humidity chamber.

3. A method according to claim 1, further comprising covering said small volume of extraction solution over said thin layer of biological sample on said flat surface of said supporting base, to prevent evaporation of said extraction solution and condensation of water vapor into said extraction solution.

4. A method according to claim 1, wherein said biological sample is a paraffin-embedded tissue sample, and wherein a step of de-paraffinization is inserted between step a and step b.

5. A method according to claim 1, wherein the following step is inserted between step a and step b:

placing a surface of a solid object close to said thin layer of biological sample to form a capillary space between said surface of said solid object and said flat surface of said supporting base.

6. A method according to claim 1, wherein the following step is inserted between step b and step c:

placing a surface of a solid object close to said thin layer of biological sample to form a capillary space between said surface of said solid object and said flat surface of said supporting base.

7. A method according to claim 1, wherein said supporting base is a transparent supporting base that facilitates microscopic examination.

8. A method according to claim 7, further comprising the following two steps inserted between steps a and b:

identifying an area of interest on said thin layer of biological sample under a microscope, and, removing unwanted area(s) on said thin layer of biological sample from said transparent supporting base.

9. A method according to claim 8, wherein said unwanted area(s) on said thin layer of biological sample is removed from said supporting base by scraping with an edge of a hard object.

10. A method according to claim 8, wherein said transparent supporting base is a flat sheet made of glass or synthetic polymers, and wherein said unwanted area(s) on said thin layer of biological sample is removed from said supporting base by cutting out said area of interest on said thin layer of biological sample attached to said transparent supporting base.

11. A method according to claim 10, wherein said area of interest on said thin layer of biological sample attached to said transparent supporting base is contacted by said small volume of extraction buffer in a container.

12. A method according to claim 1, further comprising applying ultrasound to said small volume of extraction solution during incubation.

13. A method according to claim 1, further comprising subjecting said small volume of extraction solution collected after incubation to downstream solution-based analysis.

14. A method according to claim 1, further comprising subjecting said thin layer of said biological sample remaining on said flat surface of supporting base to slide-based analysis after incubation and collection of said extraction solution.

15. A method for extracting biological molecules from a thin layer of biological sample attaching to a flat surface of a supporting base, comprising:

a) contacting said thin layer of said biological sample with a small volume of extraction solution, said extraction solution facilitates dissolution of predetermined biological molecules from said biological sample,

b) incubating at a temperature for a length of time, and,

d) collecting said extraction solution which contains said predetermined biological molecules extracted from said biological sample.

16. A method according to claim 15, wherein incubation is performed in a humidity chamber.

17. A method according to claim 15, further comprising covering said small volume of extraction solution over said thin layer of biological sample on said flat surface of supporting base, to prevent evaporation of said extraction solution and condensation of water vapor into said extraction solution.

18. A method according to claim 15, wherein said thin layer of biological sample is a paraffin-embedded tissue section, and wherein a step of de-paraffinization is inserted before step a.

19. A method according to claim 15, wherein the following step is inserted before step a:

placing a surface of a solid object close to said thin layer of biological sample to form a capillary space between said surface of said solid object and said flat surface of said supporting base.

20. A method according to claim 15, wherein the following step is inserted between step a and step b:

placing a surface of a solid object close to said thin layer of biological sample to form a capillary space between said surface of said solid object and said flat surface of said supporting base.

21. A method according to claim 15, wherein said supporting base is a transparent supporting base that facilitates microscopic examination.

22. A method according to claim 21, further comprising the following two steps inserted before steps a:

identifying an area(s) of interest on said thin layer of biological sample under a microscope, and, removing unwanted area(s) on said thin layer of biological sample from said transparent supporting base.

23. A method according to claim 22, wherein said unwanted area(s) on said thin layer of biological sample is removed from said supporting base by scraping with an edge of a hard object.

24. A method according to claim 22, wherein said transparent supporting base is a flat sheet made of glass or synthetic polymers, and wherein said unwanted area(s) on said thin layer of biological sample is removed from said transparent supporting base by cutting out said area(s) of interest on said thin layer of biological sample attaching to said transparent supporting base.

25. A method according to claim 24, wherein said area of interest on said thin layer of biological sample attaching to said transparent supporting base is contacted by said small volume of extraction buffer in a container.

26. A method according to claim 15, further comprising subjecting said small volume of extraction solution collected after incubation to downstream solution-based analysis.

27. A method according to claim 15, further comprising applying ultrasound to said small volume of extraction solution during incubation.

28. A method according to claim 15, further comprising subjecting said thin layer of said biological sample remaining on said flat surface of supporting base to slide-based analysis after incubation and collection of said extraction solution.