Cresyl Violet Staining of Tissue Sections: A Staining Procedure to Visualize Cartilage and Bones in Frozen Mouse Embryo Sections

Prepare three labeled 50-milliliter centrifuge tubes. Add 45 milliliters of 80% ethanol to each tube. Place three tubes on ice. Place a tube containing 45 milliliters of 95% ethanol and a tube containing 100% ethanol on ice. Next, add 45 milliliters of xylene to a 50-milliliter centrifuge tube at room temperature.

Thaw a PEN membrane slide briefly by placing it against a gloved hand, and wash it twice for 30 seconds in each of the tubes containing 80% ethanol with agitation, to remove the optimal cutting temperature compound. Then, lay the slide on a sheet of aluminum foil. Pipette 0.8 milliliters of 0.1% of cresyl violet in 50% ethanol onto the slide, and stain for 30 seconds.

Wash the slide in the third tube containing 80% ethanol for 30 seconds. Dehydrate the slide by passing it through the tube containing 95% ethanol for 30 seconds, the tube containing 100% ethanol for 30 seconds, and finally the tube containing xylene for 30 seconds. After this, place the slides on a delicate task wiper for 5 minutes to let the xylene drain and allow the sections to dry.