2-9. Lee's Methylene Blue -- Basic Fuchin (Ref 1)
Staining Procedures For Plastic Embedded Tissue
Verified at the Applications Laboratory of the Biomedical Division, Sorvall
Microtomes
SOLUTIONS:
| Methylene Blue Stock |
|
0.5g |
|
Methylene blue |
|
400.0ml |
|
Distilled water |
|
|
|
|
| Basic Fuchsin Stoc |
|
0.5g |
|
Basic fuchsin |
|
400.0ml |
|
Distilled water |
|
|
|
|
| Stock Buffe |
|
|
|
Stock buffer pH 6.8 to 7.2. Higher pH levels produce bluer
results. Lower pH values give pinker results. |
|
|
|
|
| Working Solutio |
|
12.0ml |
|
Methylene blue solution |
|
12.0ml |
|
Basic fuchsin solution |
|
21.0ml |
|
Stock buffer |
|
15.0ml |
|
Ethyl alcohol, 95%
Filter |
|
|
|
|
STAINING PROCEDURE:
- Stain in working solution for 5-15 seconds or longer.
- Rinse in distilled water.
- Dip in 95% alcohol to remove background stain, 1-2 dips.
Blow dry.
Mount.
RESULTS:
Nuclei -- blue; cytoplasm, mitochondria, cilia, certain cell granules
--shades of red and pink; cartilage and most cell granules -- blue
to purple.
Warning:Some of the chemicals
used for the staining procedures given in this section may be hazardous
if misused. For this reason, read and observe all warnings and cautions
provided by the manufacturer for each chemical before proceeding
with a staining procedure.
Note: In order to prevent sections from
loosening from the slides during staining, all sections should
be heat-fixed (60ºC to
100ºC) to the slides for a minimum of 2-5 minutes prior to staining,
preferably at the time the sections are mounted on the slides. |
