Kluver-Barrera Method for Myelin and Nerve Cells
EMS Catalog #: 26681
Fixation:
Formalin, 10% Buffered Neutral (#15740)
Sections:
Paraffin @ 15 to 20 microns
Staining:
- Deparaffinize and hydrate to 95% alcohol.
- Leave in Luxol Fast Blue Solution (#26681-01) in 56°C oven overnight.
- Rinse off excess stain with 95% alcohol.
- Rinse in distilled water.
- Differentiate the slides singly in Lithium Carbonate Solution (#26681-04) for 30 seconds.
- Continue differentiation in 70% alcohol until the gray matter is clear and white matter sharply defined.
- Check microscopically. Repeat the differentiation if necessary starting at step 5.
- When differentiation is complete, place in distilled water.
- When all slides have been collected in distilled water, add fresh distilled water.
- Counterstain in (Cresyl Violet Acetate),Working solution for 6 minutes.
- To prepare working solution add 15 drops of Acetic Acid, 10% (#26681-03) to 100 ml of
Cresyl Violet Acetate, 0.1% (#26681-02) just before use. Filter.
- Rinse in 2 changes of 95% alcohol.
- Continue the dehydration through two changes of absolute ethyl alcohol and xylene, two changes each, for 2 minutes.
- Mount with resinous medium.
Results:
Myelin, including phospholipids |
Blue to Green cells |
Cells and cell products |
Pink to violet |
- NOTE: The Luxol Fast Blue procedure works well when combined with other procedures such as the Bodian Method.
References:
Kluver, H., and Barrera, E. : A method for the combined staining of cells and fibers in the Nervous system. J. Nueropath. Exp. Neurol. 12:400-403, 1953.
AFIP, Lab Methods in Histothechnology, ed. Edna B. Prophet, Bob Mills. Jacquelyn B. Arrmyl, Leslie H. Sobin, M.D. Wash, D.C. American Registry of Pathology, c. 1992, p. 94-95.
Product Information
Kluver-Barrera Method for Myelin and Nerve Cells