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<DIV><FONT face=Arial size=2>I wonder if someone can assist me in solving this
problem.I retrieved a hematoxylin and eosin/phloxine stained slide from 1998 and
discovered it was faded.The tissue section was mounted in a coverslipping
machine that uses coverslipping tape composed of cellulose triacetate and is
coated on one side with resinous mountant.This is the procedure I followed to
decoverslip,decolorise and restain: I placed the slide in acetone for 7
minutes to remove the coverslip and then put the slide in xylene for 6
hours to remove any residual resinous mountant.I took the slide through
graded ethanols to water after which I decolorised with acid/alcohol for one
minute, took the slide back to water and restained the nuclei with Gill's
Hematoxylin ( for 5 min. ) and counterstained with a mixture of 2%
eosin y and 1% phloxine B with 4 drops of conc. glacial acetic acid added (
for 3 min. ).The nuclear staining turned out pale and the counter stain was
patchy almost as if something was blocking the tissue's ability to retain the
dyes I cannot figure out why the tissue is not staining with the same intensity
as it did in 1998.Please keep in mind the slide was left out in natural light
for a while which resulted in it becoming faded.</FONT></DIV>
<DIV><FONT face=Arial
size=2>.....................................................<BR>Peter
Smale<BR>Anatomical Pathology<BR>National Health Laboratory Service<BR>Chris
Hani Baragwanath Hospital<BR>Soweto, Johannesburg, South
Africa<BR>tel: +2711 4898711<BR>fax: +2711 4898717 <BR>e-mail:
<A href="mailto:peter.smale@nhls.ac.za">peter.smale@nhls.ac.za</A></FONT></DIV>
<DIV> </DIV>
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