Compositions comprising nanomaterials and method for using such compositions for histochemical processes

• US 8,703,490 B2
• Filed: 06/01/2009
• Issued: 04/22/2014
• Est. Priority Date: 06/05/2008
• Status: Active Grant

• Current Assignee: Ventana Medical Systems Incorporated (Roche Holding AG)
• Original Assignee: Ventana Medical Systems Incorporated (Roche Holding AG)
• Inventors: Jackson, Merrill, Herman, Michael, Hoben, Grace, Sebastiao, Noemi, Cockayne, Scott, Ferrea, Heather
• Primary Examiner(s): COLE, MONIQUE T
• Application Number:
  US12/995,790
• Publication Number:
  US 20110091897A1
• Time in Patent Office: 1,786 Days
• Field of Search: 436/10, 436/17, 436/8, 436/18, 436/63, 435/40.5, 435/40.51, 435/40.52, 252/408.1
• US Class Current: 436/17
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: G01N 1/30 Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
• CPC Class Codes: Y10T 436/10 Composition for standardization, calibration, simulation, stabilization, preparation or preservation; processes of use in preparation for chemical testing
• CPC Class Codes: Y10T 436/101666 Particle count or volume standard or control [e.g., platelet count standards, etc.]
• CPC Class Codes: Y10T 436/107497 Preparation composition [e.g., lysing or precipitation, etc.]
• CPC Class Codes: Y10T 436/108331 Preservative, buffer, anticoagulant or diluent

Abstract

Histochemical process compositions comprising at least one nanoparticle in an amount effective to reduce or substantially eliminate the average number of spots per slide that result from a sample staining protocol are disclosed. Various nanoparticles, or combinations thereof, including metals, metal alloys, metal oxides, ceramics, functionalized metals or metalloids, and other miscellaneous nanoparticles, such as carbon nanoparticles and diamond nanoparticles, can be used. The nanoparticle concentration typically is from about 2 parts per million to about 20 parts per million. One embodiment of a disclosed method concerns applying a histochemical process composition to a sample, followed by performing a staining protocol on the sample. Particular embodiments concern dispensing a nanosolution onto a sample using an automated system, heating the sample, and performing a sample staining process.

Claims

1. A histochemical process composition, comprising deionized water, Tris base, boric acid, EDTA disodium dehydrate, a composition comprising 5-chloro-2-methyl-4-isothiazolin-3-one 2.30%, 2-methyl-4-isothiazolin-3-one 0.70%, modified glycol 93-95%, alkyl carboxylate 2-3%, and greater than 0 to about 25 parts per million of at least one nanoparticle.

4. The composition according to claim 1 where the nanoparticle is a metal or metal alloy.

5. The composition according to claim 4 where the nanoparticle is aluminum, copper, copper-zinc alloy, gold, gold colloid solution, a nickel-molybdenum-chromium alloy comprising approximately 55% Ni, 15-17% Mo, 14.5-16.5% Cr, 4-7% Fe and 3-4.5%, iron, iron-nickel alloy, molybdenum, nickel, palladium, platinum, silicon, silver, silver dispersion, silver-copper alloy, silver-platinum alloy, silver-tin alloy, tantalum, tin, titanium, tungsten, zinc, or combinations thereof.

6. The composition according to claim 1 where the nanoparticle is a metal oxide.

7. The composition according to claim 6 where the nanoparticle is aluminum cerium oxide, aluminum oxide, antimony tin oxide, antimony (III) oxide, barium strontium titanium oxide, barium zirconate, bismuth cobalt zinc oxide, bismuth (III) oxide, calcium oxide, cerium (IV) oxide, cerium (IV)-zirconium (IV) oxide, chromium (III) oxide, cobalt aluminum oxide, cobalt (II, III) oxide, copper aluminum oxide, copper iron oxide, copper zinc iron oxide, copper (I) oxide, copper (II) oxide, dysprosium (III) oxide, erbium (III) oxide, europium (III) oxide, gadolinium (III) oxide, holmium (III) oxide, indium tin oxide, indium (III) hydroxide, indium (III) oxide, iron nickel oxide, iron (II, III) oxide, iron (III) oxide, lanthanum (III) oxide, magnesium hydroxide, magnesium oxide, magnetic Iron oxide, neodymium (III) oxide, nickel chromium oxide, nickel cobalt, oxide, nickel zinc, iron oxide, nickel (II) oxide, praseodymium (III, IV) oxide, samarium strontium cobalt oxide, samarium(III) oxide, silica, silicon dioxide, alumina doped, terbium (III, IV) oxide, tin (IV) oxide, titanium silicon oxide, titanium (IV) oxide, titanium (IV) oxide, anatase, titanium (IV) oxide, mixture of rutile and anatase, titanium (IV) oxide, rutile, tungsten (VI) oxide, ytterbium (III) oxide, yttrium aluminum oxide, yttrium iron oxide, yttrium (III) oxide, zinc iron oxide, zinc oxide, zirconium (IV) oxide, zirconium(IV) oxide-yttria stabilized, or combinations thereof.

8. The composition according to claim 6 where the nanoparticle is alumina (Al₂O₃), silica (SiO₂), titania (TiO₂), or combinations thereof.

9. The composition according to claim 1 where the nanoparticle is a ceramic.

10. The composition according to claim 9 where the nanoparticle is aluminum nitride, aluminum titanate, barium ferrite, barium titanate (IV), amorphous calcium phosphate, calcium titanate, calcium zirconate, hydroxyapatite, magnesium aluminate, silicon carbide, silicon nitride, strontium ferrite, strontium titanate, titanium carbonitride, titanium carbonitride (7:

3), titanium (IV) carbide, zinc titanate, zirconium (IV) silicate, or combinations thereof.

11. The composition according to claim 1 where the nanoparticle is a functionalized metal or metalloid.

12. The composition according to claim 11 where the nanoparticle is (1-mercaptoundec-11-yl)tetra(ethylene glycol) functionalized gold nanoparticles solution, 1-mercapto-(triethylene glycol) methyl ether functionalized gold nanoparticles solution, 3-aminopropyl functionalized silica, 3-aminopropyl-(3-oxobutanoic acid) functionalized silica, decanethiol functionalized silver nanoparticles solution, dodecanethiol functionalized gold nanoparticles solution, dodecanethiol functionalized silver nanoparticles solution, octanethiol functionalized gold nanoparticles solution, and combinations thereof.

13. The composition according to claim 1 where the nanoparticle is a carbon nanoparticle, a diamond nanoparticle, or combinations thereof.

14. The composition according to claim 1 where the nanoparticle has a concentration of from about 2 parts per million to about 20 parts per million.

15. The composition according to claim 1 where the nanoparticle has a concentration of from about 2.5 parts per million to about 15 parts per million.

2. A histochemical process composition, comprising deionized water, sodium citrate dihydrate, citric acid monohydrate, polyoxyethylenesorbitan monolaurate, a composition comprising 5-chloro-2-methyl-4-isothiazolin-3-one 2.30%, 2-methyl-4-isothiazolin-3-one 0.70%, modified glycol 93-95%, alkyl carboxylate 2-3%, and greater than 0 to about 25 parts per million of at least one nanoparticle.

16. The composition according to claim 2 where the nanoparticle is a metal or metal alloy.

17. The composition according to claim 16 where the nanoparticle is aluminum, copper, copper-zinc alloy, gold, gold colloid solution, a nickel-molybdenum-chromium alloy comprising approximately 55% Ni, 15-17% Mo, 14.5-16.5% Cr, 4-7% Fe and 3-4.5%, iron, iron-nickel alloy, molybdenum, nickel, palladium, platinum, silicon, silver, silver dispersion, silver-copper alloy, silver-platinum alloy, silver-tin alloy, tantalum, tin, titanium, tungsten, zinc, or combinations thereof.

18. The composition according to claim 2 where the nanoparticle is a metal oxide.

19. The composition according to claim 18 where the nanoparticle is aluminum cerium oxide, aluminum oxide, antimony tin oxide, antimony (III) oxide, barium strontium titanium oxide, barium zirconate, bismuth cobalt zinc oxide, bismuth (III) oxide, calcium oxide, cerium (IV) oxide, cerium (IV)-zirconium (IV) oxide, chromium (III) oxide, cobalt aluminum oxide, cobalt (II, III) oxide, copper aluminum oxide, copper iron oxide, copper zinc iron oxide, copper (I) oxide, copper (II) oxide, dysprosium (III) oxide, erbium (III) oxide, europium (III) oxide, gadolinium (III) oxide, holmium (III) oxide, indium tin oxide, indium (III) hydroxide, indium (III) oxide, iron nickel oxide, iron (II, III) oxide, iron (III) oxide, lanthanum (III) oxide, magnesium hydroxide, magnesium oxide, magnetic Iron oxide, neodymium (III) oxide, nickel chromium oxide, nickel cobalt, oxide, nickel zinc, iron oxide, nickel (II) oxide, praseodymium (III, IV) oxide, samarium strontium cobalt oxide, samarium(III) oxide, silica, silicon dioxide, alumina doped, terbium (III, IV) oxide, tin (IV) oxide, titanium silicon oxide, titanium (IV) oxide, titanium (IV) oxide, anatase, titanium (IV) oxide, mixture of rutile and anatase, titanium (IV) oxide, rutile, tungsten (VI) oxide, ytterbium (III) oxide, yttrium aluminum oxide, yttrium iron oxide, yttrium (III) oxide, zinc iron oxide, zinc oxide, zirconium (IV) oxide, zirconium(IV) oxide-yttria stabilized, or combinations thereof.

20. The composition according to claim 18 where the nanoparticle is alumina (Al₂O₃), silica (SiO₂), titania (TiO₂), or combinations thereof.

21. The composition according to claim 2 where the nanoparticle is a ceramic.

22. The composition according to claim 21 where the nanoparticle is aluminum nitride, aluminum titanate, barium ferrite, barium titanate (IV), amorphous calcium phosphate, calcium titanate, calcium zirconate, hydroxyapatite, magnesium aluminate, silicon carbide, silicon nitride, strontium ferrite, strontium titanate, titanium carbonitride, titanium carbonitride (7:

3), titanium (IV) carbide, zinc titanate, zirconium (IV) silicate, or combinations thereof.

23. The composition according to claim 2 where the nanoparticle is a functionalized metal or metalloid.

24. The composition according to claim 23 where the nanoparticle is (1-mercaptoundec-11-yl)tetra(ethylene glycol) functionalized gold nanoparticles solution, 1-mercapto-(triethylene glycol) methyl ether functionalized gold nanoparticles solution, 3-aminopropyl functionalized silica, 3-aminopropyl-(3-oxobutanoic acid) functionalized silica, decanethiol functionalized silver nanoparticles solution, dodecanethiol functionalized gold nanoparticles solution, dodecanethiol functionalized silver nanoparticles solution, octanethiol functionalized gold nanoparticles solution, and combinations thereof.

25. The composition according to claim 2 where the nanoparticle is a carbon nanoparticle, a diamond nanoparticle, or combinations thereof.

26. The composition according to claim 2 where the nanoparticle has a concentration of from about 2 parts per million to about 20 parts per million.

27. The composition according to claim 2 where the nanoparticle has a concentration of from about 2.5 parts per million to about 15 parts per million.

3. A histochemical process composition, comprising deionized water, sodium citrate dihydrate, citric acid monohydrate, 5-chloro-2-methyl-4-isothiazolin-3-one 2.30%, 2-methyl-4-isothiazolin-3-one 0.70%, modified glycol 93-95%, alkyl carboxylate 2-3%, and greater than 0 to about 25 parts per million of at least one nanoparticle.

28. The composition according to claim 3 where the nanoparticle is a metal or metal alloy.

29. The composition according to claim 28 where the nanoparticle is aluminum, copper, copper-zinc alloy, gold, gold colloid solution, a nickel-molybdenum-chromium alloy comprising approximately 55% Ni, 15-17% Mo, 14.5-16.5% Cr, 4-7% Fe and 3-4.5%, iron, iron-nickel alloy, molybdenum, nickel, palladium, platinum, silicon, silver, silver dispersion, silver-copper alloy, silver-platinum alloy, silver-tin alloy, tantalum, tin, titanium, tungsten, zinc, or combinations thereof.

30. The composition according to claim 3 where the nanoparticle is a metal oxide.

31. The composition according to claim 30 where the nanoparticle is aluminum cerium oxide, aluminum oxide, antimony tin oxide, antimony (III) oxide, barium strontium titanium oxide, barium zirconate, bismuth cobalt zinc oxide, bismuth (III) oxide, calcium oxide, cerium (IV) oxide, cerium (IV)-zirconium (IV) oxide, chromium (III) oxide, cobalt aluminum oxide, cobalt (II, III) oxide, copper aluminum oxide, copper iron oxide, copper zinc iron oxide, copper (I) oxide, copper (II) oxide, dysprosium (III) oxide, erbium (III) oxide, europium (III) oxide, gadolinium (III) oxide, holmium (III) oxide, indium tin oxide, indium (III) hydroxide, indium (III) oxide, iron nickel oxide, iron (II, III) oxide, iron (III) oxide, lanthanum (III) oxide, magnesium hydroxide, magnesium oxide, magnetic Iron oxide, neodymium (III) oxide, nickel chromium oxide, nickel cobalt, oxide, nickel zinc, iron oxide, nickel (II) oxide, praseodymium (III, IV) oxide, samarium strontium cobalt oxide, samarium(III) oxide, silica, silicon dioxide, alumina doped, terbium (III, IV) oxide, tin (IV) oxide, titanium silicon oxide, titanium (IV) oxide, titanium (IV) oxide, anatase, titanium (IV) oxide, mixture of rutile and anatase, titanium (IV) oxide, rutile, tungsten (VI) oxide, ytterbium (III) oxide, yttrium aluminum oxide, yttrium iron oxide, yttrium (III) oxide, zinc iron oxide, zinc oxide, zirconium (IV) oxide, zirconium(IV) oxide-yttria stabilized, or combinations thereof.

32. The composition according to claim 30 where the nanoparticle is alumina (Al₂O₃), silica (SiO₂), titania (TiO₂), or combinations thereof.

33. The composition according to claim 3 where the nanoparticle is a ceramic.

34. The composition according to claim 33 where the nanoparticle is aluminum nitride, aluminum titanate, barium ferrite, barium titanate (IV), amorphous calcium phosphate, calcium titanate, calcium zirconate, hydroxyapatite, magnesium aluminate, silicon carbide, silicon nitride, strontium ferrite, strontium titanate, titanium carbonitride, titanium carbonitride (7:

3), titanium (IV) carbide, zinc titanate, zirconium (IV) silicate, or combinations thereof.

35. The composition according to claim 3 where the nanoparticle is a functionalized metal or metalloid.

36. The composition according to claim 35 where the nanoparticle is (1-mercaptoundec-11-yl)tetra(ethylene glycol) functionalized gold nanoparticles solution, 1-mercapto-(triethylene glycol) methyl ether functionalized gold nanoparticles solution, 3-aminopropyl functionalized silica, 3-aminopropyl-(3-oxobutanoic acid) functionalized silica, decanethiol functionalized silver nanoparticles solution, dodecanethiol functionalized gold nanoparticles solution, dodecanethiol functionalized silver nanoparticles solution, octanethiol functionalized gold nanoparticles solution, and combinations thereof.

37. The composition according to claim 3 where the nanoparticle is a carbon nanoparticle, a diamond nanoparticle, or combinations thereof.

38. The composition according to claim 3 where the nanoparticle has a concentration of from about 2 parts per million to about 20 parts per million.

39. The composition according to claim 3 where the nanoparticle has a concentration of from about 2.5 parts per million to about 15 parts per million.

40. A method, comprising:

providing a biological sample;

applying a histochemical process composition to the sample, the composition comprising deionized water, Tris base, boric acid, EDTA disodium dehydrate, a composition comprising 5-chloro-2-methyl-4-isothiazolin-3-one 2.30%, 2-methyl-4-isothiazolin-3-one 0.70%, modified glycol 93-95%, alkyl carboxylate 2-3%, and greater than 0 to about 25 parts per million of at least one nanoparticle; and

performing a histochemical process on the sample.

41. The method according to claim 40 where performing a histochemical process comprises performing cell conditioning on the sample using a cell conditioning composition.

42. The method according to claim 40 where performing a histochemical process includes heating the biological sample and histochemical process composition to a temperature of from about 60°

to about 101°

C.

43. The method according to claim 40 where the nanoparticle is selected from aluminum, copper, copper-zinc alloy, gold, gold colloid solution, a nickel-molybdenum-chromium alloy comprising approximately 55% Ni, 15-17% Mo, 14.5-16.5% Cr, 4-7% Fe and 3-4.5%, iron, iron-nickel alloy, molybdenum, nickel, palladium, platinum, silicon, silver, silver dispersion, silver-copper alloy, silver-platinum alloy, silver-tin alloy, tantalum, tin, titanium, tungsten, zinc, aluminum cerium oxide, aluminum oxide, antimony tin oxide, antimony (III) oxide, barium strontium titanium oxide, barium zirconate, bismuth cobalt zinc oxide, bismuth (III) oxide, calcium oxide, cerium (IV) oxide, cerium (IV)-zirconium (IV) oxide, chromium (III) oxide, cobalt aluminum oxide, cobalt (II, III) oxide, copper aluminum oxide, copper iron oxide, copper zinc iron oxide, copper (I) oxide, copper (II) oxide, dysprosium (III) oxide, erbium (III) oxide, europium (III) oxide, gadolinium (III) oxide, holmium (III) oxide, indium tin oxide, indium (III) hydroxide, indium (III) oxide, iron nickel oxide, iron (II, III) oxide, iron (III) oxide, lanthanum (III) oxide, magnesium hydroxide, magnesium oxide, magnetic Iron oxide, neodymium (III) oxide, nickel chromium oxide, nickel cobalt, oxide, nickel zinc, iron oxide, nickel (II) oxide, praseodymium (III, IV) oxide, samarium strontium cobalt oxide, samarium(III) oxide, silica, silicon dioxide, alumina doped, terbium (III, IV) oxide, tin (IV) oxide, titanium silicon oxide, titanium (IV) oxide, titanium (IV) oxide, anatase, titanium (IV) oxide, mixture of rutile and anatase, titanium (IV) oxide, rutile, tungsten (VI) oxide, ytterbium (III) oxide, yttrium aluminum oxide, yttrium iron oxide, yttrium (III) oxide, zinc iron oxide, zinc oxide, zirconium (IV) oxide, zirconium(IV) oxide-yttria stabilized, aluminum nitride, aluminum titanate, barium ferrite, barium titanate (IV), amorphous calcium phosphate, calcium titanate, calcium zirconate, hydroxyapatite, magnesium aluminate, silicon carbide, silicon nitride, strontium ferrite, strontium titanate, titanium carbonitride, titanium carbonitride (7:

3), titanium (IV) carbide, zinc titanate, zirconium (IV) silicate, (1-mercaptoundec-11-yl)tetra(ethylene glycol) functionalized gold nanoparticles solution, 1-mercapto-(triethylene glycol) methyl ether functionalized gold nanoparticles solution, 3-aminopropyl functionalized silica, 3-aminopropyl-(3-oxobutanoic acid) functionalized silica, decanethiol functionalized silver nanoparticles solution, dodecanethiol functionalized gold nanoparticles solution, dodecanethiol functionalized silver nanoparticles solution, octanethiol functionalized gold nanoparticles solution, a carbon nanoparticle, a diamond nanoparticle, or combinations thereof.

44. The method according to claim 40 where the composition comprises a nanoparticle selected from alumina (Al₂O₃), silica (SiO₂), titania (TiO₂), or combinations thereof.

45. The method according to claim 40 where the composition has a nanoparticle concentration of from about 2 parts per million to about 20 parts per million.

46. The method according to claim 40 where the composition has a nanoparticle concentration of from about 2.5 parts per million to about 15 parts per million.

47. The method according to claim 40 further comprising performing at least one additional histochemical processing step.

48. A method, comprising:

providing a biological sample;

applying a histochemical process composition to the sample, the composition comprising deionized water, sodium citrate dihydrate, citric acid monohydrate, polyoxyethylenesorbitan monolaurate, a composition comprising 5-chloro-2-methyl-4-isothiazolin-3-one 2.30%, 2-methyl-4-isothiazolin-3-one 0.70%, modified glycol 93-95%, alkyl carboxylate 2-3%, and greater than 0 to about 25 parts per million of at least one nanoparticle; and

performing a histochemical process on the sample.

49. The method according to claim 48 where performing a histochemical process comprises performing cell conditioning on the sample using a cell conditioning composition.

50. The method according to claim 48 where performing a histochemical process includes heating the biological sample and histochemical process composition to a temperature of from about 60°

to about 101°

C.

51. The method according to claim 48 where the nanoparticle is selected from aluminum, copper, copper-zinc alloy, gold, gold colloid solution, a nickel-molybdenum-chromium alloy comprising approximately 55% Ni, 15-17% Mo, 14.5-16.5% Cr, 4-7% Fe and 3-4.5%, iron, iron-nickel alloy, molybdenum, nickel, palladium, platinum, silicon, silver, silver dispersion, silver-copper alloy, silver-platinum alloy, silver-tin alloy, tantalum, tin, titanium, tungsten, zinc, aluminum cerium oxide, aluminum oxide, antimony tin oxide, antimony (III) oxide, barium strontium titanium oxide, barium zirconate, bismuth cobalt zinc oxide, bismuth (III) oxide, calcium oxide, cerium (IV) oxide, cerium (IV)-zirconium (IV) oxide, chromium (III) oxide, cobalt aluminum oxide, cobalt (II, III) oxide, copper aluminum oxide, copper iron oxide, copper zinc iron oxide, copper (I) oxide, copper (II) oxide, dysprosium (III) oxide, erbium (III) oxide, europium (III) oxide, gadolinium (III) oxide, holmium (III) oxide, indium tin oxide, indium (III) hydroxide, indium (III) oxide, iron nickel oxide, iron (II, III) oxide, iron (III) oxide, lanthanum (III) oxide, magnesium hydroxide, magnesium oxide, magnetic Iron oxide, neodymium (III) oxide, nickel chromium oxide, nickel cobalt, oxide, nickel zinc, iron oxide, nickel (II) oxide, praseodymium (III, IV) oxide, samarium strontium cobalt oxide, samarium(III) oxide, silica, silicon dioxide, alumina doped, terbium (III, IV) oxide, tin (IV) oxide, titanium silicon oxide, titanium (IV) oxide, titanium (IV) oxide, anatase, titanium (IV) oxide, mixture of rutile and anatase, titanium (IV) oxide, rutile, tungsten (VI) oxide, ytterbium (III) oxide, yttrium aluminum oxide, yttrium iron oxide, yttrium (III) oxide, zinc iron oxide, zinc oxide, zirconium (IV) oxide, zirconium(IV) oxide-yttria stabilized, aluminum nitride, aluminum titanate, barium ferrite, barium titanate (IV), amorphous calcium phosphate, calcium titanate, calcium zirconate, hydroxyapatite, magnesium aluminate, silicon carbide, silicon nitride, strontium ferrite, strontium titanate, titanium carbonitride, titanium carbonitride (7:

3), titanium (IV) carbide, zinc titanate, zirconium (IV) silicate, (1-mercaptoundec-11-yl)tetra(ethylene glycol) functionalized gold nanoparticles solution, 1-mercapto-(triethylene glycol) methyl ether functionalized gold nanoparticles solution, 3-aminopropyl functionalized silica, 3-aminopropyl-(3-oxobutanoic acid) functionalized silica, decanethiol functionalized silver nanoparticles solution, dodecanethiol functionalized gold nanoparticles solution, dodecanethiol functionalized silver nanoparticles solution, octanethiol functionalized gold nanoparticles solution, a carbon nanoparticle, a diamond nanoparticle, or combinations thereof.

52. The method according to claim 48 where the composition comprises a nanoparticle selected from alumina (Al₂O₃), silica (SiO₂), titania (TiO₂), or combinations thereof.

53. The method according to claim 48 where the composition has a nanoparticle concentration of from about 2 parts per million to about 20 parts per million.

54. The method according to claim 48 where the composition has a nanoparticle concentration of from about 2.5 parts per million to about 15 parts per million.

55. The method according to claim 48 further comprising performing at least one additional histochemical processing step.

56. A method, comprising:

providing a biological sample;

applying a histochemical process composition to the sample, the composition comprising deionized water, sodium citrate dihydrate, citric acid monohydrate, 5-chloro-2-methyl-4-isothiazolin-3-one 2.30%, 2-methyl-4-isothiazolin-3-one 0.70%, modified glycol 93-95%, alkyl carboxylate 2-3%, and greater than 0 to about 25 parts per million of at least one nanoparticle; and

performing a histochemical process on the sample.

57. The method according to claim 56 where performing a histochemical process comprises performing cell conditioning on the sample using a cell conditioning composition.

58. The method according to claim 56 where performing a histochemical process includes heating the biological sample and histochemical process composition to a temperature of from about 60°

to about 101°

C.

59. The method according to claim 56 where the nanoparticle is selected from aluminum, copper, copper-zinc alloy, gold, gold colloid solution, a nickel-molybdenum-chromium alloy comprising approximately 55% Ni, 15-17% Mo, 14.5-16.5% Cr, 4-7% Fe and 3-4.5%, iron, iron-nickel alloy, molybdenum, nickel, palladium, platinum, silicon, silver, silver dispersion, silver-copper alloy, silver-platinum alloy, silver-tin alloy, tantalum, tin, titanium, tungsten, zinc, aluminum cerium oxide, aluminum oxide, antimony tin oxide, antimony (III) oxide, barium strontium titanium oxide, barium zirconate, bismuth cobalt zinc oxide, bismuth (III) oxide, calcium oxide, cerium (IV) oxide, cerium (IV)-zirconium (IV) oxide, chromium (III) oxide, cobalt aluminum oxide, cobalt (II, III) oxide, copper aluminum oxide, copper iron oxide, copper zinc iron oxide, copper (I) oxide, copper (II) oxide, dysprosium (III) oxide, erbium (III) oxide, europium (III) oxide, gadolinium (III) oxide, holmium (III) oxide, indium tin oxide, indium (III) hydroxide, indium (III) oxide, iron nickel oxide, iron (II, III) oxide, iron (III) oxide, lanthanum (III) oxide, magnesium hydroxide, magnesium oxide, magnetic Iron oxide, neodymium (III) oxide, nickel chromium oxide, nickel cobalt, oxide, nickel zinc, iron oxide, nickel (II) oxide, praseodymium (III, IV) oxide, samarium strontium cobalt oxide, samarium(III) oxide, silica, silicon dioxide, alumina doped, terbium (III, IV) oxide, tin (IV) oxide, titanium silicon oxide, titanium (IV) oxide, titanium (IV) oxide, anatase, titanium (IV) oxide, mixture of rutile and anatase, titanium (IV) oxide, rutile, tungsten (VI) oxide, ytterbium (III) oxide, yttrium aluminum oxide, yttrium iron oxide, yttrium (III) oxide, zinc iron oxide, zinc oxide, zirconium (IV) oxide, zirconium(IV) oxide-yttria stabilized, aluminum nitride, aluminum titanate, barium ferrite, barium titanate (IV), amorphous calcium phosphate, calcium titanate, calcium zirconate, hydroxyapatite, magnesium aluminate, silicon carbide, silicon nitride, strontium ferrite, strontium titanate, titanium carbonitride, titanium carbonitride (7:

3), titanium (IV) carbide, zinc titanate, zirconium (IV) silicate, (1-mercaptoundec-11-yl)tetra(ethylene glycol) functionalized gold nanoparticles solution, 1-mercapto-(triethylene glycol) methyl ether functionalized gold nanoparticles solution, 3-aminopropyl functionalized silica, 3-aminopropyl-(3-oxobutanoic acid) functionalized silica, decanethiol functionalized silver nanoparticles solution, dodecanethiol functionalized gold nanoparticles solution, dodecanethiol functionalized silver nanoparticles solution, octanethiol functionalized gold nanoparticles solution, a carbon nanoparticle, a diamond nanoparticle, or combinations thereof.

60. The method according to claim 56 where the composition comprises a nanoparticle selected from alumina (Al₂O₃), silica (SiO₂), titania (TiO₂), or combinations thereof.

61. The method according to claim 56 where the composition has a nanoparticle concentration of from about 2 parts per million to about 20 parts per million.

62. The method according to claim 56 where the composition has a nanoparticle concentration of from about 2.5 parts per million to about 15 parts per million.

63. The method according to claim 56 further comprising performing at least one additional histochemical processing step.