Calibration slides for digital pathology
In some embodiments, the present disclosure is directed to coatings or thin films comprising a dye or stain embedded within a matrix, e.g. a polymer matrix.
1. A calibration system comprising:
(a) an optically transparent substrate;
(b) a film having a substantially uniform thickness disposed on at least a portion of the optically transparent substrate, the film comprising a chromogenic precipitate embedded within a polymer, wherein the chromogenic precipitate is uniformly dispersed within the polymer.
2. The calibration system of claim 1 , wherein the polymer is a hydrogel.
3. The calibration system of claim 2 , wherein the hydrogel comprises crosslinked gelatin.
4. The calibration system of claim 3 , wherein the crosslinked gelatin is derived from gelatin and an aldehyde.
5. The calibration system of claim 2 , wherein the hydrogel is derived from a gelling agent and a crosslinking agent.
6. The calibration system of claim 1 , wherein the polymer is an acrylate.
7. The calibration system of claim 1 , wherein the chromogenic precipitate is a reaction product of a chromogen and an enzyme.
8. The calibration system of claim 7 , wherein the chromogen is selected from the group consisting of 5-Bromo-4-Chloro-3-Indolyl Phosphate; 4-Chloro-2-methyl benzenediazonium; 3,3′-Diaminobenzidine; 3,3′,5,5;-tetramethylbenzidine; 4-chloro-1-naphthol; 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid); o-phenylenediamine dihydrochloride; p-Nitrophenyl Phosphate; 5-Bromo-4-Chloro-3-indolyl-B-D-Galactopyranoside; and 2,2,5-5-tetra-p-nitrophenyl-3,3-(3,3-dimethoxy-4,4-biphenylene)-di tetrazolium chloride.
9. The calibration system of claim 8 , wherein the chromogen is 3,3′-Diaminobenzidine.
10. The calibration system of claim 8 , wherein the enzyme is selected from the group consisting of alkaline phosphatase, horseradish peroxidase, glycosylases, and glucose oxidase.
11. The calibration system of claim 1 , wherein a ratio of the chromogenic precipitate to the polymer ranges from about 0.1 to about 50.
12. A set of slides comprising:
(a) a first slide having a first coating disposed on at least a portion of an upper surface of the first slide, the first coating comprising a stain present in a polymeric matrix at a first concentration;
(b) a second slide having a second coating disposed on at least a portion of an upper surface of the second slide, the second coating comprising the stain present in the polymeric matrix at a second concentration;
wherein the first concentration and the second concentration are different.
13. The set of slides of claim 12 , wherein the stain is a chromogenic precipitate.
14. The set of slides of claim 13 , wherein the chromogenic precipitate is derived from one of 5-Bromo-4-Chloro-3-Indolyl Phosphate; 4-Chloro-2-methyl benzenediazonium; 3,3′-Diaminobenzidine; 3,3′,5,5;-tetramethylbenzidine; 4-chloro-1-naphthol; 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid); o-phenylenediamine dihydrochloride; p-Nitrophenyl Phosphate; 5-Bromo-4-Chloro-3-indolyl-B-D-Galactopyranoside; or 2,2,5-5-tetra-p-nitrophenyl-3,3-(3,3-dim ethoxy-4,4-biphenylene)-di tetrazolium chloride.
15. The set of slides of claim 12 , wherein the second concentration is at least twice the first concentration.
16. A kit comprising:
(i) a first set of coating reagents comprising:
a. a first intermediate coating solution comprising a first matrix forming component and an enzyme; and
b. a second intermediate coating solution comprising a chromogen, wherein the chromogen is present in the second intermediate coating solution at a first concentration; and
(ii) a second set of coating reagents comprising
a. a third intermediate coating solution comprising the first matrix forming component and the enzyme; and
b. a fourth intermediate coating solution comprising the chromogen, wherein the chromogen is present in the fourth intermediate coating solution at a second concentration.
17. The kit of claim 16 , further comprising a fifth intermediate coating solution comprising a crosslinking reagent.
18. The kit of claim 16 , wherein the chromogen is selected from the group consisting of 5-Bromo-4-Chloro-3-Indolyl Phosphate; 4-Chloro-2-methyl benzenediazonium; 3,3′-Diaminobenzidine; 3,3′,5,5;-tetramethylbenzidine; 4-chloro-1-naphthol; 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid); o-phenylenediamine dihydrochloride; p-Nitrophenyl Phosphate; 5-Bromo-4-Chloro-3-indolyl-B-D-Galactopyranoside; and 2,2,5-5-tetra-p-nitrophenyl-3,3-(3,3-dimethoxy-4,4-biphenylene)-di tetrazolium chloride.
19. The kit of claim 16 , wherein the chromogen is 3,3′-Diaminobenzidine.
20. The kit of claim 18 , wherein the enzyme is selected from the group consisting of alkaline phosphatase, horseradish peroxidase, glycosylases, and glucose oxidase.