Re: Cover flourescence ?
"Dianne Holmes (by way of Histonet)" wrote:
> We are using Rhodamine microspheres to label rat brains. Got great
> labelling - - coverslipped with DPX and lost half of the signals!!!
> HELP! Is it the xylene? Any suggestions ...
Xylene and DPX do not abolish fluorescence of rhodamine-labelled
proteins, which are denatured (made permanently insoluble) when a
apecimen passes through a chemically unreactive fixative (alcohol
etc). You are probably correct in your assumption that a solvent
dissolved the stuff the microspheres were made of. Do you know
what it was? If you don't, why did you use it? Even if there is
trade secrecy about the microspheres, their maker must surely
issue some information about solvents to avoid.
John A. Kiernan
Department of Anatomy and Cell Biology
The University of Western Ontario
London, Canada N6A 5C1
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