Kurnick Stain
for Plasma Cells
Materials
Stock pyronin
Material | Amount | |
---|---|---|
Pyronin Y | 2 | g |
Distilled water | 100 | mL |
Stock methyl green
Material | Amount | |
---|---|---|
Methyl green | 2 | g |
Distilled water | 100 | mL |
Working solution
Material | Amount | |
---|---|---|
Stock pyronin | 12.5 | mL |
Stock methyl green | 7.5 | mL |
Distilled water | 30 | mL |
Storing stock solutions
The pyronin stock solution should be extracted with chloroform ten times to ensure purity. The methyl green stock solution should be extracted a minimum of six times, until the chloroform is no longer colored with any crystal violet. Store both solutions over a layer of chloroform.
Tissue Sample
Most fixatives should be satisfactory if fixation time is not extended, but avoid potassium dichromate or picric acid. Overnight with a 10% formalin variant should be satisfactory.
Protocol
- Bring sections to distilled water via xylene and ethanol.
- Place into the staining solution for 6 minutes in a coplin jar.
- Blot gently.
- Dehydrate with n-butanol, two changes of 5 minutes each.
- Clear with xylene and mount with a resinous medium.
Expected Results
- Nuclei – green to blue-green
- Plasma cell cytoplasm, Nissl substance, nucleoli – red
- Other cell cytoplasm – pink to unstained
Notes
- As with other methyl green-pyronin methods used for demonstrating nucleic acids, in critical applications a control section should be treated with a 0.1% solution of ribonuclease for one hour at 37°C to remove RNA, and another treated with distilled water alone. Both control sections should be compared with an untreated but stained section.
Safety Note
Prior to handling any chemical, consult the Safety Data Sheet (SDS) for proper handling and safety precautions.
References
- Disbrey, B. D., (1970)
Histological laboratory methods. pp.187-188.
E. & S. Livingstone, Edinburgh and London, UK.