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Techniques. Protocols

HARRIS HEMATOXYLIN

Hematoxylin
Hematoxylin

Harris hematoxylin is a suitable dye to stain cell nuclei. It can be used as progressive and regressive staining. Harris hematoxylin is widely used for detection of cancer cells. Initially, the solution contained mercury oxide, but It can be substituted by other oxidizing substance. The oxidation transforms hematoxylin in hematein, which is the real dye.

Procedure

With mercury

5 g hematoxylin (C.I. 75290)

50 ml 100º ethanol

100 g aluminium ammonium sulfate

2,5 g mercury oxide (HgO)

Add distilled water to get 1000 ml of total solution

No mercury

5 g hematoxylin (C.I. 75290)

50 ml 100º ethanol

100 g aluminium ammonium sulfate

0,37 sodium iodate

1000 ml distilled water

Preparation

1. Dissolve hematoxylin in ethanol (it may be necessary to dilute hematoxylin in a bit of distilled water first).

2. Dissolve the aluminium ammonium sulfate in warmed distilled water.

3. Mix the two solutions made above and boil the solution.

4. Cool the solution and add the mercury oxide or sodium iodate. Mix slowly and gently.

5. If sodium iodate was added, then boil the solution again.

6. Cool the solution by placing the flask on ice or cold water

7. After cooling, filter the solution and keep in bottle protected from light.

The solution is stable for several months. To check the solution before use, put a drop of the solution on a filter paper. There should be a brown color in the center and a purple color in the periphery of the extended drop. If this is not happening, the solution may be discarded.

Products

Hematoxylin (C.I. 75290)

100º ethanol

Mercury oxide (HgO)

Sodium iodate (NaIO3)

Aluminium ammonium sulfate (AlNH4(SO4)2)

Distilled water

Labware

Test tube

Balance

Magnetic stirrer

Heating plate

Flasks

Bottle

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