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Cytopathology of the uterine cervix - digital atlas

Specimen processing - Lab technique  Rechercher Specimen processing - Lab technique

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Harris haematoxylin:
Proportions for approximately 2 liters:
  • Haematoxylin: 9 g
  • Ethanol (absolute): 90 ml
  • Potassium alun: 180 g
  • Distilled water: 1800 ml
  • Mercuric oxide: 4,5 g
  • Thymol: 9 g
Instructions:
  • Dissolve 1 g of haematoxylin in 10 ml of absolute ethanol.
  • Dissolve 20 g of potassium alun in 200 ml distilled water other gentle heat.
  • Before boiling point, add the disloved haematoxylin to the Alun solution (careful of splashing).
  • Boil the mixture as quickly as possible for few minutes.
  • Slowly add 0,5 g of mercuric oxide (take great care against splashing).
  • Quickly cool the mixing in a cold water bath.
  • Filter after 24 hours.
  • Add 1 g of thymol for conservation purposes.
Orange G:
Proportions for approximately 800 ml:
  • Orange G: 4 g
  • Distilled water: 36 ml
  • 95░ ethanol: 760 ml
  • Phosphotungstic acid: 0,12 g
Instructions:
  • Dissolve 2 g of orange G in 18 ml distilled water, under gentle heat.
  • After cooling, add 380 ml of 95░ ethanol.
  • Filter.
  • Add 0,06 g of phosphotungstic acid.
EA 50:
Proportions for approximately 800 ml :
  • Yellow eosin: 2 g
  • Light green: 2 g
  • Bismarck brown: 0,25g
  • Distilled water: 38,25 ml
  • 95░ ethanol: 1042,5 ml
  • Phosphotungstic acid: 1,8 g
  • Lithium carbonate
Instructions:
  • Dissolve 2 g of yellow eosin in 18 ml distilled water, and add 380 ml of 95░ ethanol.
  • Dissolve 2 g of light green in 18 ml distilled water, and add 380 ml of 95░ ethanol.
  • Dissolve 0,25 g of Bismarck brown in 2,25 ml distilled water, and add 147,5 ml of 95░ ethanol.
  • Take 398 ml of the light green solution, 398 ml of the eosin solution, 87,5 ml of the brown Bismarck solution.
  • Filter the mixture (the 3 sstains together).
  • Finally add 1,8 g phosphotungstic acid, 9 drops of a saturated lithium carbonate solution, and 135 ml of 95░ ethanol.
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