Surgipath SelecTech
Hematoxylin & Eosin

SelecTech 560 Hemalum   Compounding procedure
Hematoxylin 6 g Dissolve the alum and aluminum sulphate in 350 mL water.
Ammonium alum 30 g Dissolve the hematoxylin in 300 mL water.
Aluminum sulphate 50 g Dissolve the sodium iodate in 50 mL water.
Distilled water 700 mL Combine the hematoxylin and alum solutions.
Ethylene glycol 300 mL Add the sodium iodate solution.
Sodium iodate 1 g Add hydrochloric acid. Leave overnight.
Hydrochloric acid 2 mL Add the ethylene glycol, mix and filter.

 

Eosin 515   Eosin Trichrome 515
Ethanol 725 mL Ethanol 725 mL
iso-Propanol 40 mL iso-Propanol 40 mL
Methanol 36.5 mL Methanol 36.5 mL
Acetic acid 50 mL Acetic acid 50 mL
Distilled water 150 mL Distilled water 150 mL
Eosin Y, ws 2.5 g Eosin Y, ws 2.5 g
      Phloxine B 1 g
      Orange G 1 g

 

Blue Buffer   Define
Distilled water 1 L Distilled water 1 L
Trizma base 10 g Citric acid 20 g
      Sodium citrate, monobasic 20 g

 

Expected results
The procedure was designed to be used with routinely fixed and paraffin processed tissues.

Method

  1. Bring sections to water with xylene and ethanol.
  2. Place into the staining solution for the appropriate time.
  3. Rinse well with water.
  4. Differentiate with citric acid buffer.
  5. Rinse with water and blue with Trizma base solution.
  6. Rinse well with water.
  7. Counterstain with Surgipath Eosin 515 or Eosin 515 Trichrome.
  8. Dehydrate with ethanol, clear with xylene and mount with a resinous medium.

Expected results

 

Notes

  1. The hemalum should have a pH of 2-3.
  2. The Eosin 515 and Eosin Trichrome 515 should have a pH of 4-5.
  3. The Define solutions should have a pH of 3-4.
  4. The Blue buffer should have a pH of 8-8.3.
  5. As the hemalum contains hydrochloric acid it may remove small calcium deposits. These are sometimes an indicator of disease, in malignant breast lumps, for example.
  6. Although the Blue buffer, Define and the two eosin solutions are designed to be used together, Other solutions may be substituted.

 

Reference
Surgipath web site
http://surgipath.com

 


 

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