Fixative and staining solutions
The formulations, systems, and methods disclosed herein permit automated preparation of specimens (e.g., biological specimens) for examination. The disclosed formulations, systems, and methods provide fast, efficient, and highly uniform specimen processing using minimal quantities of fluids. The methods include at least a fixing phase for fixing a specimen to a substrate such as a microscope slide, a staining phase for staining the specimen, and a rinsing phase for rinsing the specimen. One or more of the fixing, staining, and rinsing phases include one or more agitation phases for distributing reagents evenly and uniformly across the specimen. The systems can be implemented as a standalone device or as a component in a larger system for preparing and examining specimens.
1. A cytological fixative solution, consisting of
a single cytological dye in an amount of from 0.5 g/L to 5.0 g/L;
0.5 mL/L to 5.0 mL/L polysorbate 20;
5 mL/L to 50 mL/L ethylene glycol;
0.10 g/L to 10 g/L HEPES sodium salt; and
alcohol.
2. The cytological fixative solution of claim 1 , wherein the polysorbate 20 is present in an amount of from 0.05% to 0.3% by volume.
3. The cytological fixative solution of claim 1 , wherein a 1:10 dilution of the solution in water has a pH of from 6 to 8 and HEPES is present in an amount of from 0.5 mM to 10 mM.
4. The cytological fixative solution of claim 1 , wherein a 1:10 dilution of the solution in water has a pH of from 6.7 to 7.3 and HEPES is present in an amount of from 0.5 mM to 10 mM.
5. The cytological fixative solution of claim 1 , wherein a 1:1000 dilution of the fixative solution in water has a UV absorbance of from 0.1 to one at a peak wavelength of from 640 to 650 nm.
6. The cytological fixative solution of claim 1 , wherein the cytological dye is methylene blue, eosin, or a fluorescein derivative.
7. The cytological fixative solution of claim 1 , wherein the alcohol is methanol, ethanol, or isopropanol.
8. The cytological fixative solution of claim 1 , wherein the cytological dye is Azure B.
9. The cytological fixative solution of claim 1 , wherein the cytological dye is a thiazine dye.