Re: Tetracycline Labeling of Bone Marrrow Biopsies
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/
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"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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