At 09:20 AM 9/3/98 +0800, you wrote:
>Dear All,
>
>I have been working with JC-1 attempting to stain the mitochondria in
>hippocampal neurons. For some reason, I cannot get a strong enough signal
>from JC-1. I have calibrated the confocal and have checked everything I can
>think of, so I am confident that it isn't the machine that is causing my
>problems. I have also compared my dye concentrations to those used by
>others who have worked with JC-1, and my solutions and methods appear to be
>ok. The only thing I can think of at this stage is that the dye may
>possibly be too old, but this is difficult for me to judge, since I have
>been unable to find out much about the stability of JC-1.
>
>I would really like to hear from anyone who has any experience with JC-1
>and who may be able to shed some light onto this problem for me.
>
>Thanks in advance for any comments and suggestions.
>
>Nici
>
>Dr. Nicolette Binz
>Australian Neuromuscular Research Institute
>4th Floor A-Block
>QE II Medical Centre
>Verdun Street
>Nedlands WA 6009
>Australia
>Ph     (61)-(08)-9346 1990
>Fax    (61)-(08)-9346 3487
>e-mail [log in to unmask]
>
We have been using many probes including JC1 to stain mitochondria in
cardiac cells. JC1 turns out to be one of the most easiest mtochondrial dye
to be used. We used 5 ug/ml JC1 and incubate the cells for 10 min at 37°C.
We then record the cells within half an hour. After that, there is a
leakage or a bleaching of the dye. I am wondering that you cannot get a
strong signal. Inddeed, you may try another batch of JC1. My experienece
with pH and Ca sensitive probes too, taught me that some probe batches from
molecular Probes have sometimes some  problems.hope 
good luck
michel
Michel Puceat, Ph.D
INSERM U390
CHU Arnaud de Villeneuve
34295 MONTPELLIER, France
Phone  lab:  (33) 4-67-41-52-44
        home: (33) 4-67-59-59-13
Fax:  (33) 4-67-41-52-42