Celestine Blue Hemalum
An Acid Resistant Nuclear Stain

Solutions

A progressive alum hematoxylin, such as Mayer's (Langeron's) or Cole's.

Proescher & Arkush' iron alum-celestine blue, or Lendrum & MacFarlane's modification of it.

Tissue sample
5µ paraffin sections of neutral buffered formalin fixed tissue are suitable. Other fixatives are likely to be satisfactory.

Method

1. Bring sections to water via xylene and ethanol.
2. Place into the iron alum-celestine blue solution for 10 minutes.
3. Wash with tap water.
4. Place into the alum hematoxylin for 10 minutes.
5. Wash with water to blue.
6. Continue with the counterstain

Expected results

• Nuclei  –  blue-black
• Other tissue  –  according to the counterstain

Notes

1. Celestine blue B may stain celloidin, obscuring the background.
2. The ferric mordanted celestine blue B attaches to DNA phosphate groups. The celestine blue B is then replaced by hematein, leaving an acid resistant, ferric mordanted nuclear stain.
3. Some workers use 5% iron alum instead of solution A, claiming similar staining
4. The celestine blue B solution was originally used without hemalum and counterstained with eosin Y. This has been suggested as a substitute for Hematoxylin and Eosin.

 

Reference
Culling, C.F.A., Allison, R.T. and Barr, W.T.
Cellular Pathology Technique, Ed.4.
Butterworth, London, UK.

Bancroft, J.D. and Stevens A. (1982)
Theory and practice of histological techniques Ed. 2
Churchill Livingstone, Edinburgh & London, UK.

Humason, G. L., (1967)
Animal tissue techniques, Ed. 2
W. H. Freeman and Company, San Francisco, Ca, USA

Drury, R.A.B. and Wallington, E.A., (1980)
Carleton's histological technique Ed. 5
Oxford University Press, Oxford, UK.

Putt, F. A.
Manual of Histopathological Staining Methods
John Wiley & Sons, New York, NY., USA