Mitochondrial Staining in oocytes

[Eleanor Kable]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Hi,

Has anyone experience in staining oocytes (particularly) pig oocytes for mitochondria??  I have a user trying to stain lipid droplets and mitochondria.
The lipid droplet staining is great using BODIPY but the mito staining is diffuse throughout the oocyte except in the lipid droplets (i.e no staining).  She is staining with mitotracker red and then fixing.

Any suggestions are welcome.

Regards,
Eleanor.



Eleanor Kable| Laboratory Manager Australian Centre for Microscopy & Microanalysis

THE UNIVERSITY OF SYDNEY
Rm 128A, Madsen Building F09 | The University of Sydney | NSW | 2006 | Australia
T [phone] | F [phone] |
E [hidden email] | W http://sydney.edu.au/acmm

Incorporating:
Australian Microscopy & Microanalysis Research Facility (AMMRF) | W http://www.ammrf.org.au
ARC Centre of Excellence for Design in Light Metals | W http://www.arclightmetals.org.au

CRICOS 00026A
This email plus any attachments to it are confidential. Any unauthorised use is strictly prohibited. If you receive this email in error, please delete it and any attachments.

[David Baddeley]

http://probes.invitrogen.com/m
*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Standard mitotracker red is not fixable

http://probes.invitrogen.com/media/pis/mp07510.pdf


some of the others claim to be, but no personal experience. Another option might be antibody labelling, e.g. against Tom20, as in http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2596623/.

cheers,
David




________________________________
 From: Eleanor Kable <[hidden email]>
To: [hidden email]
Sent: Wednesday, 22 February 2012 1:56 PM
Subject: Mitochondrial Staining in oocytes
 
*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Hi,

Has anyone experience in staining oocytes (particularly) pig oocytes for mitochondria??  I have a user trying to stain lipid droplets and mitochondria.
The lipid droplet staining is great using BODIPY but the mito staining is diffuse throughout the oocyte except in the lipid droplets (i.e no staining).  She is staining with mitotracker red and then fixing.

Any suggestions are welcome.

Regards,
Eleanor.



Eleanor Kable| Laboratory Manager Australian Centre for Microscopy & Microanalysis

THE UNIVERSITY OF SYDNEY
Rm 128A, Madsen Building F09 | The University of Sydney | NSW | 2006 | Australia
T [phone] | F [phone] |
E [hidden email] | W http://sydney.edu.au/acmm

Incorporating:
Australian Microscopy & Microanalysis Research Facility (AMMRF) | W http://www.ammrf.org.au
ARC Centre of Excellence for Design in Light Metals | W http://www.arclightmetals.org.au

CRICOS 00026A
This email plus any attachments to it are confidential. Any unauthorised use is strictly prohibited. If you receive this email in error, please delete it and any attachments.

[Iain Johnson]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

The fact that MitoTracker Red nonspecifically stains yolk platelets has
been reported several times in the literature including
http://www.ncbi.nlm.nih.gov/pubmed/11739575.  However the alternative
described in that paper (the styryl dye 4-Di-2-Asp) would not be my first
choice.  I would try MitoTracker Deep Red FM, which is structurally quite a
different dye to MitoTracker Red CMXRos, though both have the chloromethyl
functionality required for coupling to proteins and thereby fixation.

If the researcher is willing to spend the modest amount of additional time
required, an antibody (as suggested earlier) or a mitochondrially-targeted
GFP fusion will have better localization specificity than any low molecular
weight dye.

Iain

Iain Johnson Consulting
Eugene, OR
[phone]

<http://www.ncbi.nlm.nih.gov/pubmed/11739575>

On Tue, Feb 21, 2012 at 5:32 PM, David Baddeley <[hidden email]
> wrote:

> http://probes.invitrogen.com/m
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Standard mitotracker red is not fixable
>
> http://probes.invitrogen.com/media/pis/mp07510.pdf
>
>
> some of the others claim to be, but no personal experience. Another option
> might be antibody labelling, e.g. against Tom20, as in
> http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2596623/.
>
> cheers,
> David
>
>
>
>
> ________________________________
>  From: Eleanor Kable <[hidden email]>
> To: [hidden email]
> Sent: Wednesday, 22 February 2012 1:56 PM
> Subject: Mitochondrial Staining in oocytes
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Hi,
>
> Has anyone experience in staining oocytes (particularly) pig oocytes for
> mitochondria??  I have a user trying to stain lipid droplets and
> mitochondria.
> The lipid droplet staining is great using BODIPY but the mito staining is
> diffuse throughout the oocyte except in the lipid droplets (i.e no
> staining).  She is staining with mitotracker red and then fixing.
>
> Any suggestions are welcome.
>
> Regards,
> Eleanor.
>
>
>
> Eleanor Kable| Laboratory Manager Australian Centre for Microscopy &
> Microanalysis
>
> THE UNIVERSITY OF SYDNEY
> Rm 128A, Madsen Building F09 | The University of Sydney | NSW | 2006 |
> Australia
> T [phone] | F [phone] |
> E [hidden email] | W http://sydney.edu.au/acmm
>
> Incorporating:
> Australian Microscopy & Microanalysis Research Facility (AMMRF) | W
> http://www.ammrf.org.au
> ARC Centre of Excellence for Design in Light Metals | W
> http://www.arclightmetals.org.au
>
> CRICOS 00026A
> This email plus any attachments to it are confidential. Any unauthorised
> use is strictly prohibited. If you receive this email in error, please
> delete it and any attachments.
>