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Kurnick Stain for Plasma Cells

Kurnick Stain

for Plasma Cells

5
steps
3
materials

Materials

Stock pyronin

MaterialAmount
Pyronin Y2g
Distilled water100mL

Stock methyl green

MaterialAmount
Methyl green2g
Distilled water100mL

Working solution

MaterialAmount
Stock pyronin12.5mL
Stock methyl green7.5mL
Distilled water30mL

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

  1. Bring sections to distilled water via xylene and ethanol.
  2. Place into the staining solution for 6 minutes in a coplin jar.
  3. Blot gently.
  4. Dehydrate with n-butanol, two changes of 5 minutes each.
  5. 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

  1. Disbrey, B. D., (1970)
    Histological laboratory methods. pp.187-188.
    E. & S. Livingstone, Edinburgh and London, UK.