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Simon:

Commercial interest acknowledged in the following.  

The best form of DiI to use for this application would be the VyBrant
DiI labeling solution (Probes ctalog number V22885). In applications
like this, DiI sometimes has problems with non-uniform labeling,
typically manifested by a few intensely stained cells superimposed on a
much more weakly stained population. 

There are many good alternatives:

For plasma membrane specific-staining, Alexa Fluor 594 WGA conjugates
work well (Probes catalog numbers W11262 or I34406). There may be
penetration issues in a thick tissue with these.  


FM4-64 (Probes Catalog Number T3166) which is highly water soluble, is
good for quick staining/viusalization, but will get endocytosed over
time.

CellTrace BODIPY TR (Probes Catalog number C34556) is a new dye that we
developed with Mark Cooper
(http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&do
pt=Abstract&list_uids=15614774&query_hl=1)
for counterstaining GFP in zebrafish embryos.  Easy to apply - just soak
the animal in dye solution. It stains primarily intracellular membranes
rather than the PM.  Penetration is excellent without any
permeabilization and the dye can be fixed later on if you need to.

Best Regards,

Iain Johnson
Molecular Probes

-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]] On
Behalf Of Simon Walker
Sent: Tuesday, June 07, 2005 9:28 AM
To: [log in to unmask]
Subject: Membrane label for tissue

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Can anyone recommend a fluorescent dye (and suitable protocol) to label
membranes (plasma membrane)in tissue (Drosophila eye).  The tissue is
expressing GFP so something which emits red would be ideal.  I see
Molecular Probes, sorry Invitrogen, sell their lipohpilic tracers (e.g.
Dil) as crystals, oils and pastes but I'm not sure which would be best
for
this application.  Any advice gratefully received.
Thanks
Simon