CoverslippingRe: [Histonet] Fluorescein Question

[Gayle Callis]

Sara,

Try it, but keep sections in the dark to avoid photobleaching.  Fluorescein 
should survive deparaffinization and then coverslipping.  Putting in an 
extra xylene to ensure good paraffin removal before putting a coverglass on 
may be of help.  We have some who do that here but they are requesting 
thick sections ~ 10 um or so.  I generally just air dry at RT in a dark 
place.  If you want to know the effects of temperature on FITC, contact 
Molecular Probes technical services, they will know about this also.

However, if the tissues are fixed with formalin, you may be battling 
autofluorescence.  At 08:48 AM 4/13/2007, you wrote:
>A current project is cutting slides to check for fluorescein expression.
>The first time I did this, I was asked to air-dry the slides and mount
>with aqueous medium; as you can expect, the coverage was "iffy" at best
>with lifting of the coverslip, etc.  Is there any reason I could not dry
>these in a 60 degree oven as usual, then deparaffinize and coverslip out
>of xylene?  Would this process compromise the results? Help? Thanks!
>
>
>
>Sally Breeden, HT(ASCP)
>
>NM Dept. of Agriculture
>
>Veterinary Diagnostic Services
>
>PO Box 700
>
>Albuquerque, NM  87106
>
>505-841-2576
>
>
>
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Gayle Callis
MT,HT,HTL(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610