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Phenolic MGP Solutions

Phenolic MGP Solutions

7
steps
7
materials

Materials

Table 1. Comparison of phenolic methyl green –pyronin solutions for plasma cells

MaterialFormula
PappenheimUnnaSandifordScott & FrenchLangeronKurnick
Distilled water100mL82mL75mL80mL100mL100mL
GlycerolmL16mL20mL16mLmLmL
Ethanol, 100%0mL2mL0mL4mLmLmL
Ethanol, 95%mLmL5mLmLmLmL
Phenol0.25g0.4g1.5g1.6g5gg
Methyl green0.3g0.12g0.15g0.8g2g0.6g
Pyronin Y0.7g0.2g0.5g0.2g2g1g

Tissue Sample

Most fixatives should be satisfactory.

Protocol

These solutions have been included for comparison purposes, and the formulae have been adjusted to make 100 mL of each solution to facilitate the comparison. See the individual methods for details of their use and the actual formula. There is no single method for their application, but the details below may be used as a starting point.

  1. Bring sections to water via xylene and ethanol.
  2. Place in the MGP solution for 10 minutes at room temperature.
  3. Rinse with distilled water.
  4. Differentiate with absolute ethanol if required.
  5. Complete dehydration with acetone if necessary.
  6. Clear with xylene and mount with a resinous medium.

Expected Results

  • Nuclei  –  blue-green
  • Plasma cell cytoplasm  –  red
  • Other cell cytoplasm  –  pink

Notes

  • The time and temperature in step 2 may need to be increased.
  • Differentiation may not be required, depending on the time and temperature of staining.
  • Various fluids have been used for dehydration: acetone, n-butanol, t-butanol, etc.

Safety Note

Prior to handling any chemical, consult the Safety Data Sheet (SDS) for proper handling and safety precautions.

References

  1. Gray, Peter. (1954)
    The Microtomist’s Formulary and Guide. p. 355
    Originally published by: The Blakiston Co.
    Republished by: Robert E. Krieger Publishing Co.
  2. Disbrey, B. D., (1970)
    Histological laboratory methods.
    E. & S. Livingstone, Edinburgh and London, UK.