[Histonet] restaining of hematoxylin and eosin tissue sections

From:"Peter Smale"

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.
Peter Smale
Anatomical Pathology
National Health Laboratory Service
Chris Hani Baragwanath Hospital
Soweto, Johannesburg, South Africa
tel:   +2711 4898711
fax:  +2711 4898717
e-mail: peter.smale@nhls.ac.za
This message is for the designated recipient only and may contain priviliged,
prorietary, or otherwise privare information. If you have received it in error,
please notify the sender immediately and delete the original.
Any other use of the email by you is prohibited.

<< Previous Message | Next Message >>