Re: Tetracycline Labeling of Bone Marrrow Biopsies

From:"J. A. Kiernan"

The purpose of the staining isn't clear, but
tetracycline can be used in conjunction with
alum, in the same way as oxidized haematoxylin,
to provide a fluorescent nuclear stain. Here's
a method.

Staining solution

Tetracycline hydrochloride:    0.5 g
Water:                         100 ml
Aluminium potassium sulphate
  (AlK(SO4)2.12.H2O):          5.0 g

The solution is stable for about one month, 
becoming decreasingly efficacious as its colour 
changes from yellow to orange-brown.

Procedure

1. Stain hydrated sections for 5 minutes.
2. Wash in 3 changes of water.
3. Dehydrate through graded alcohols, 
   clear in xylene, and mount in a 
   non-fluorescent resinous medium.
4. Examine by fluorescence microscopy, 
   with excitation by ultraviolet or blue 
   light.

Result

  Nuclei emit a yellow to orange 
  fluorescence.
-- 
-------------------------
John A. Kiernan
Department of Anatomy and Cell Biology
The University of Western Ontario
London,   Canada   N6A 5C1
   kiernan@uwo.ca
   http://publish.uwo.ca/~jkiernan/
_________________________________________________

"Jones, Charnell E" wrote:
> 
> Hi All,
> 
> I have a pathologist that is interested in labeling bone marrow biopsies with the antibiotic tetracycline and then using fluorescence to visualize. Is anyone familiar with this technique? I found a reference for this technique in Sheehan for methacrylate embedded sections, but I wanted to know if this could be done on paraffin processed tissue? If you have any other references or procedures for this technique, I would appreciate it. Thanks in advance.
> 
> Charnell E. Jones, BS, HTL(ASCP)
> CompuNet Clinical Laboratories
> Dayton, OH
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