RE: Squamous epithelial cells in lung tissue.

From:

"Smith, Allen"

You might try zirconyl hematoxylin, which stains the same cells that alcian blue at pH 2.5 does but stains them in a violet that is less intense than "halcyon blue".

Dissolve 100 mg hematoxylin in 5 ml absolute ethanol.

Add 5 ml freshly made 0.1% aqueous sodium iodate.

Add 400 mg zirconyl chloride octahydrate.

Add 10 ml glycerol mixed with 30 ml water.

Stir 5 minutes. This staining solution keeps for 1 year.

Stain hydrated sections 10 min.

Rinse in distilled water, 2 changes, 2 min. each.

Counterstain if you like. My current preference is:

Kernechtrot, 5 min.

Running tap water, 1 min.

0.2% aqueous tropaeolin O, 5 min.

Distilled water, 2 seconds.

70% ethanol, 5 min.

Dehydrate, clear, and mount.

Reference for the stain is: Smith, A.A. (2000) Zirconyl hematoxylin: a new stain for acidic mucins. Biotechnic and Histochemistry 75: 124-131. The counterstain is mentioned in a paper just submitted to Acta Histochemica.

Allen A. Smith, Ph.D.
Barry University
School of Graduate Medical Sciences
Podiatric Medicine and Surgery
Miami Shores, Florida 33161-6695

-----Original Message-----
From: Bill Sinai [mailto:bills@icpmr.wsahs.nsw.gov.au]
Sent: Wednesday, November 13, 2002 6:36 PM
To: histonet
Subject: Squamous epithelial cells in lung tissue.

Dear All,

For the last twenty years we have been using Harold Attwood's Method for Amniotic Fluid Embolism. Now we find that Alcian green is unavailable. In the past we were able to make do with a mix of Alcian Blue and Alcian Yellow at pH 2.5. Now we find that mixing these two dyes simply causes them to form a gel like mixture which does not stain (almost colloidal).

Does anyone have any idea of where I might be able to obtain Alcian Green or a dye we can use as a substitute in this stain.

Alcian Blue or Alcian Yellow alone is too strong a colour to allow the squamous cells to be seen as they are usually very pale pink.

Any advice would be appreciated.

Many thanks

Bill Sinai
Laboratory Manager
Tissue Pathology
ICPMR
P.O. Box 533
Wentworthville NSW 2145
Ph 02 9845 7774

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