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Leder Esterase for Mast Cells

Leder Esterase

for Mast Cells

7
steps
12
materials

Materials

Pararosanilin

MaterialAmount
Pararosanilin0.5g
Distilled water20mL
Hydrochloric acid, conc.2.5mL

Warm gently and filter. Refrigerate.

Nitrosylated pararosanilin

MaterialAmount
Pararosanilin solution0.1mL
Sodium nitrite, 4% aqueous0.1mL

The sodium nitrite solution must be fresh. After mixing, let stand for one minute. Use immediately.

Sorenson Stock A

MaterialAmount
Disodium hydrogen phosphate2.73g
Distilled water250mL

Sorenson Stock B

MaterialAmount
Potassium dihydrogen phosphate2.27g
Distilled water250mL

Sorenson working buffer

MaterialAmount
Sorenson stock A41mL
Sorenson stock B9mL

Incubating medium

MaterialAmount
Naphthol-ASD chloroacetate10mg
N,N-dimethyformamide1mL
Sorenson’s working buffer35mL
Nitrosylated pararosanilin0.2mL

Combine in the order given. Mix well, filter and use immediately.

Light green

MaterialAmount
Light green SF yellowish1g
Distilled water100mL
Glacial acetic acid1mL

Tissue Sample

4µ paraffin sections of formalin fixed tissue are suitable.

Protocol

  1. Bring sections to distilled water.
  2. Place in the incubating medium for 40 minutes.
  3. Wash in running tap water for 5 minutes.
  4. Counterstain with light green for 30 seconds.
  5. Wash in running tap water for 5 min.
  6. Air dry.
  7. Clear in xylol and mount.

Expected Results

  • Esterase activity (mast cells)  –  red
  • Background  –  green

Notes

  • An alternative to air drying, which avoids some of the artifact introduced, is to blot the section, then wash with xylene. This is repeated until the sections are cleared.

Safety Note

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

References

  1. Leder, L.D. (1964)
    Klinische Wochenschrift