One of my project this semester was to improve the Cresyl Violet (CV) staining protocol used in the laboratory. Cresyl Violet is a dye used to stain brain tissues and it is commonly used in histological sectioning. The dye is basic and therefore it binds to ribosomes that are RNA rich and nuclei of cells, which are acidic. Also, the dye leaves a purple color on the stained neuronal tissues. For the purpose of most of our studies, Cresyl Violet staining is used to analyze the lesion size and location in the brain of mice. This staining is important in our research because the slides need to be stained properly in order for us to get accurate results. Our research team had been noticing that the slides that were stained with CV were different shades of purple and some slides had no color on them. This indicated that there was an error because the slides should all look very similar. Since I had some background information on staining from microbiology laboratory, I started to observe that some slides were left in ethanol for longer periods of times. Ethanol is used to dehydrate the slides and that’s why some slides were over-decolorized. Research shows that there are a variety of different protocols for Cresyl Violet staining.1 The indicated time that the slides …show more content…
The 2% Potassium Ferrocyanide solution was made using 2.5 grams of Potassium Ferrocyanide and 125.0 milliliters of distilled water. The 2% Hydrochloric acid solution was created using 2.5 milliliters of Hydrochloric Acid and 122.5 milliliters of distilled water. The Potassium Ferrocyanide-Hydrochloric Acid Solution was prepared by mixing 125.0 milliliters of 2% Potassium Ferrocyanide and 125.0 milliliters of 2% Hydrochloric Acid. The Mix equal parts of 2% hydrochloric acid and 2% potassium ferrocyanide solution JUST before