Histological Staining Techniques
Luxol fast blue/cresyl violet
Kluver and Barrera, 1954.
Use 15µm sections.
Take to 95% alcohol.
Stain in luxol fast blue solution - 2 hours at 60oC* or preferably
at 37oC. (Use a screw-topped plastic coplin jar.)
Wash in 95% alcohol.
Rinse in distilled.
Start differentiation by agitating for 10 seconds in a jar of 0.005% lithium
carbonate. This solution begins to loose its differentiating ability after
a few uses so replace it often with fresh.
Differentiate for 30-60 seconds in 70% alcohol.
Rinse all slides in distilled water and check the staining with a microscope.
Repeat the three steps above until grey and white matter are clearly distinguishable.
Nuclei should be colourless and myelin should stand out turquoise on a
very pale grey/blue background.
Stain in 0.1% cresyl violet in 1% acetic acid - 10 minutes.
Wash in distilled.
Rinse in 70% alcohol.
Differentiate in 70% alcohol until only nuclei and nissl substance are
purple - approx. 30 seconds.
Rinse quickly in 100% alcohol, clear and mount.
Myelin - blue.
Red blood cells - turquoise.
Nuclei and nissl substance - purple.
* For the 2 hour timing use 0.05% lithium carbonate to differentiate
instead of 0.005%, but be careful, it works much more quickly!
Luxol fast blue solution
Luxol fast blue MBS - 1g
95% alcohol - 1 litre
10% acetic acid - 5mls
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