Toluidine Blue Staining of Peripheral Nerve Sections: A Technique to Stain Myelinated Nerve Fibers For Peripheral Nerve Structure Visualization

To stain the nerve sections, use a plastic pipette or micro pipettor to add a drop of toluidine blue solution on the top of the nerve sections. After 20 to 30 seconds, rinse off all toluidine blue solution excess, by gently dipping the slides into a jar of deionized water, repeating 3 to 4 times until sections are clear.

Dry the slides for at least 15 minutes at 60 degrees Celsius, or overnight at room temperature. After drying, add regular mounting medium, and cover the sections with a coverslip. Finally, examine the mounted sections under a light microscope. A 100X oil immersion lens is recommended for detailed images to calculate g-ratios.