RV: MitoTracker Red and Fixation
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Konstantín Levitskiy |
RV: MitoTracker Red and Fixation
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Hi all. Sorry. It takes me a
little. First of all it was in 2002. Some parts of the HD with the images were
corrupted. Finally here is the post of images. https://postimg.org/gallery/24sbw7vr0/b946c360/ The first image is the
plasmid EYFP-mito expressed in a fibroblast or in a smooth muscle cell (in vivo). The last 6 are the in
vivo images. The rest of them are some fixed cells with PFA or in vivo with loss of colocalization after the
exposure to Antimycine A. Best
regards, Dr. Konstantín Levitskiy Servicio de Microscopía InstitutodeBiomedicinadeSevilla - IBiS Campus
del Hospital Universitario Virgen del Rocío Avda.
Manuel Siurot s/nº 41013
Sevilla Tlfno:
955 92 3030 Email:
[hidden email] Web:
www.ibis-sevilla.es AVISO
SOBRE CONFIDENCIALIDAD. Este mensaje y sus
anexos son confidenciales y pueden estar protegidos por disposiciones legales.
Si Vd. no es el destinatario del mismo, por favor, notifíquenoslo inmediatamente
y destruya o devuelva el original. No deberá copiar este mensaje ni sus anexos
o usarlo para propósito alguno, ni divulgar su contenido a ninguna persona. P El medio ambiente es nuestra responsabilidad. Por
favor, antes de imprimir este mensaje, compruebe que sea necesario hacerlo. De: Indig, Fred
(NIH/NIA/IRP) [E] [mailto:[hidden email]] Dear Dr. Levitskiy, Your posting is very
interesting. Would you be willing to share an image or two illustrating the
problem? Sincerely,
From: Microscopia-IBIS
[[hidden email]] ***** To join, leave or search the
confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com
and include the link in your posting. ***** Hi. About 10 years ago we
bought MitoTrackerDeepRed (MTDR) and we had some use in vivo. When we fixed we
noticed something different. We expressed pEYFP-Mito in cells and after
checking total colocalization with a dye we washed with a PFA, seeing how the
dye was internalized in another place but with an aspect “similar to the
mitochondria”. No was colocalization at all after fixing. We asked about
it to Molecular probes, but with no answer. Regards, Dr. Konstantín Levitskiy Servicio de Microscopía InstitutodeBiomedicinadeSevilla - IBiS Campus
del Hospital Universitario Virgen del Rocío Avda.
Manuel Siurot s/nº 41013
Sevilla Tlfno:
955 92 3030 Email:
[hidden email] Web:
www.ibis-sevilla.es AVISO
SOBRE CONFIDENCIALIDAD. Este mensaje y sus
anexos son confidenciales y pueden estar protegidos por disposiciones legales.
Si Vd. no es el destinatario del mismo, por favor, notifíquenoslo
inmediatamente y destruya o devuelva el original. No deberá copiar este mensaje
ni sus anexos o usarlo para propósito alguno, ni divulgar su contenido a
ninguna persona. P El medio ambiente es nuestra responsabilidad. Por
favor, antes de imprimir este mensaje, compruebe que sea necesario hacerlo. De: Confocal Microscopy List [mailto:[hidden email]]
En nombre de Iain Johnson Aldehyde fixation will certainly cause destaining
because it kills the mitochondrial membrane potential that drives the
electrophoretic uptake of the dye. Any dye that isn't coupled to protein will
wash out. Iain Sent from my iPhone
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