ELF-staining in double in situ whole mounts.

Michael Lardelli mlardelli at genetics.adelaide.edu.au
Tue Mar 2 18:52:13 EST 1999

Hi out there,

I am writing to see whether anyone can give me advice on staining using the
ELF-97 Endogenous Phosphatase Detection Kit from Molecular Probes. We have
tried using it twice to visualise transcripts in double in situ
hybridisation in whole embryos and we seem to see the patterns that we
should. However the nature of the signal is very difficult to interpret.
Rather than seeing a fluorescent signal from a whole cell, we seem to get
what I call "hairy crystals" - thin, curved slivers of rather intense
illumination that are very difficult to colocalise with other markers under
normal fluorescence microscopy. The density of the "hairy crystals" is not
great enough to give confidence that every cell that could be staining is
staining - hence it is not very useful to us. Has anyone seen this before?
Is there a solution?

Best wishes,


Michael Lardelli
Department of Genetics
University of Adelaide
South Australia, 5005

Tel.    (Aust. = 61) (0)8 8303 3212  direct
        (Aust. = 61) (0)8 8303 5633  departmental secretary
Fax.    (Aust. = 61) (0)8 8303 4399
e-mail. mlardelli at genetics.adelaide.edu.au

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