Dear Dr. Carvalho

An alternative tehcnique to anti-FITC antibodies in electron microcopy may be
the photoconversion of DAB by the irradiation of the FITC-marked cells using a
laser or a fluorescent microscope.
I never used this technique but it appears to be relatively easy.
This technique is described by Pagano and Chen in Annals of the New York Academy
of Siences, 845:152-160, 1998.

I hope it works
Good Luck!

            [log in to unmask]
Francisco Javier Hernandez-Blazquez
IARC-WHO
Unit of Multistage Carcinogenesis
150 Cours Albert Thomas
Lyon - France

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Tina Carvalho wrote:

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> Hi, all-
>
> A researcher has approached me with confocal images of part of an insect's
> nervous system that has cells that particularly light up with FITC.
> They don't know what these cells are, and they want to be able to locate
> these cells in the TEM.  They are looking for something in particular, but
> I am not allowed to be more specific.
>
> Their idea is to have HRP-conjugated anti-FITC, hoping for a dark reaction
> product that I will be able to find in the TEM.
>
> I haven't done any HRP staining in (yikes!) decades, and so do not have a
> protocol handy.  Could someone suggest one or, perhaps better, an
> alternative plan?  I am very open to suggestions!  Since all they know
> about these cells is that they fluoresce with FITC, I haven't come up with
> any other labelling ideas.  When I asked if they could microinject the
> cells, I got blank and then fearful stares.
>
> Thanks in advance!
>
> Mahalo,
> Tina
>
> ****************************************************************************
> * Tina (Weatherby) Carvalho               * [log in to unmask]           *
> * Biological Electron Microscope Facility * (808) 956-6251                 *
> * University of Hawaii at Manoa           * http://www.pbrc.hawaii.edu/bemf*
> ****************************************************************************