[Histonet] restaining of hematoxylin and eosin tissue sections

Peter Smale peter.smale <@t> nhls.ac.za
Wed Nov 26 01:13:07 CST 2003


To Juan Gutierrez from San Antonio TX
Thanks for your help.What you said makes perfect sense.You see, when we
bought this coverslipper, the company that sold it to us didn't give us all
the info on how to decoverslip, except that the slides must be put in
acetone first.

Regards
.....................................................
Peter Smale
Anatomical Pathology
National Health Laboratory Service
Chris Hani Baragwanath Hospital
Soweto, Johannesburg
tel:   +2711 4898711
fax:  +2711 4898717
e-mail: peter.smale <@t> nhls.ac.za

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----- Original Message -----
From: "GUTIERREZ, JUAN" <juan.gutierrez <@t> christushealth.org>
To: "Peter Smale" <peter.smale <@t> nhls.ac.za>;
<histonet <@t> lists.utsouthwestern.edu>
Sent: Tuesday, November 25, 2003 6:21 PM
Subject: RE: [Histonet] restaining of hematoxylin and eosin tissue sections


> You never got rid of the mountant.  The mountant used on the tape is
> activated when it comes in contact with xylene. So by putting the slides
> back in xylene, you only reactivated it.  After you remove the
> coverslip, you must alternately rinse in water and dip in fresh acetone(
> not the one you used to take the covers. off).  The endpoint of this
> step is easy to see because the mountant will turn white when it comes
> in contact with the water.  Keep dipping and rinsing until all the white
> gunk is gone.  Good luck!
>
> Juan C. Gutierrez, HT(ASCP)
> Histology Supervisor
> Christus Santa Rosa Healthcare
> San Antonio, TX  78207
> (210)704-2533
>
> -----Original Message-----
> From: Peter Smale [mailto:peter.smale <@t> nhls.ac.za]
> Sent: Tue 11/25/2003 2:17 AM
> To: histonet <@t> lists.utsouthwestern.edu
> Cc:
> Subject: [Histonet] restaining of hematoxylin and eosin tissue sections
>
>
> 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 <@t> nhls.ac.za <mailto:peter.smale <@t> 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.
> -----------------------------------------------------
>
>





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