Nissl Staining
Steps
Must have six containers
Add distilled water to first container.
Add 70% alcohol to second container.
Add absolute alcohol to third container.
Add clear safe to fourth container.
Add Cresyl violet stain to fifth container.
Take slides, make sure backside has glass, and place into first container.
Leave them there for 2 minutes before moving onto the next step.
Take out slides and place them into the second container. Leave them in there for 2-4 minutes (4 minutes preferably)
Take out slides and place them into the third container--this finishes the dehydration process. Leave them in there for 5 minutes.
Move the slides into the clear safe. Make sure no "smoke appears." If it shows, move back to alcohol.
Pull slides out of clear safe and move to absolute alcohol for 4 minutes.
Move into 70% alcohol for 4 minutes.
Move into water for 2 minutes.
Place slides into stain for 20 minutes.
Add acid alcohol into 6th container.
Place acid in-between water and 70% alcohol.
After 20 minutes, move slides into water; refill water as necessary (after color change).
Move into acid alcohol; refill as needed.Â
Move from acid alcohol into 70% alcohol.
Move into absolute alcohol for a little longer.
Move into clear safe.
Prepare cover slips by gripping edges and separating them.
Use one-sided q-tip to apply paramount to slides.
Remove slide from clear safe.
Have the slide brain side up, facing away from you.
Place paramount on bottom of slide facing away from you.
Take a cover slip and place it perpendicular to slide, and move until facing slide.
After this, the staining is done.