new prolong gold antifade with dapi

[Pamela Young]

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Hello Listers,

I have a user who just repeated an experiment for the millionth time but using the new formula of Prolong Gold Antifade with Dapi (P36935).  This new formula can be kept at room temp, but her former prolong had to be stored in the freezer.  As far as we can figure, the new prolong is the only thing changed in her protocol.  And unfortunately, instead of imaging beautiful FITC-labeled elastin, she saw green elastin and bright green cytosol.  She previously had seen in the FITC channel nothing from cells and green fibers (elastin) around the cells.

Has anyone tried the new prolong gold?  Because that is the only thing we can think of that might be different.  We are going to test by repeating and just mounting in PBS.

Thanks,
Pam

Dr Pamela A. Young | Light and Optical Microscopist
Australian Centre for Microscopy & Microanalysis

THE UNIVERSITY OF SYDNEY
Rm 116A, Madsen Building F09 | The University of Sydney | NSW | 2006 | Australia
T [phone] | F [phone]
E [hidden email]<mailto:[hidden email]> | W http://sydney.edu.au/acmm

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

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[Feinstein, Timothy]

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Hi Pam,

From their website it looks like Mol Probes (no commercial interest) only
recommends their new Prolong Gold for Alexa dyes.  For other stuff
including ³traditional dyes" they recommend a different stuff called
Prolong Diamond.  

https://www.lifetechnologies.com/us/en/home/life-science/cell-analysis/cell
ular-imaging/fluorescence-microscopy-and-immunofluorescence-if/mounting-med
ium-antifades/prolong-gold-antifade.html

I would guess this has to do with their custom antifade, similar to how
phenylenediamine plays very badly with Cy dyes.

Cheers,


TF

Timothy Feinstein, Ph.D. | Confocal Manager
333 Bostwick Ave., N.E., Grand Rapids, Michigan 49503
Phone: [phone] | Email: [hidden email]







On 7/17/14, 3:59 AM, "Pamela Young" <[hidden email]> wrote:

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>
>Hello Listers,
>
>I have a user who just repeated an experiment for the millionth time but
>using the new formula of Prolong Gold Antifade with Dapi (P36935).  This
>new formula can be kept at room temp, but her former prolong had to be
>stored in the freezer.  As far as we can figure, the new prolong is the
>only thing changed in her protocol.  And unfortunately, instead of
>imaging beautiful FITC-labeled elastin, she saw green elastin and bright
>green cytosol.  She previously had seen in the FITC channel nothing from
>cells and green fibers (elastin) around the cells.
>
>Has anyone tried the new prolong gold?  Because that is the only thing we
>can think of that might be different.  We are going to test by repeating
>and just mounting in PBS.
>
>Thanks,
>Pam
>
>Dr Pamela A. Young | Light and Optical Microscopist
>Australian Centre for Microscopy & Microanalysis
>
>THE UNIVERSITY OF SYDNEY
>Rm 116A, Madsen Building F09 | The University of Sydney | NSW | 2006 |
>Australia
>T [phone] | F [phone]
>E [hidden email]<mailto:[hidden email]> | W
>http://scanmail.trustwave.com/?c=129&d=v4LH048qAI0z3gfJsKATBR2BxFxR5VhNMAv
>HSAQbnw&u=http%3a%2f%2fsydney%2eedu%2eau%2facmm
>
>Incorporating:
>Australian Microscopy & Microanalysis Research Facility (AMMRF) | W
>http://scanmail.trustwave.com/?c=129&d=v4LH048qAI0z3gfJsKATBR2BxFxR5VhNMFm
>SSgAVnQ&u=http%3a%2f%2fwww%2eammrf%2eorg%2eau<http://scanmail.trustwave.co
>m/?c=129&d=v4LH048qAI0z3gfJsKATBR2BxFxR5VhNMAzGHAIVlA&u=http%3a%2f%2fwww%2
>eammrf%2eorg%2eau%2f>
>ARC Centre of Excellence for Design in Light Metals | W
>http://scanmail.trustwave.com/?c=129&d=v4LH048qAI0z3gfJsKATBR2BxFxR5VhNMFv
>BTlQczA&u=http%3a%2f%2fwww%2earclightmetals%2eorg%2eau<http://scanmail.tru
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>
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>use is strictly prohibited. If you receive this email in error, please
>delete it and any attachments.

[jerie]

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Dear Pamela,

I found DAPI concentrations too high by a factor of at least 10.000 in
the old Prolong, in particular when working with weak other labels
(why are you using FITC?, is this labeled primary AB? consider to use
Alexa anti FITC 2ndary to boost the signal), high DAPI conc produces a
strong background, in particular in the green.


Cheers, Jens

On 7/17/14, Pamela Young <[hidden email]> wrote:

> *****
> 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.
> *****
>
> Hello Listers,
>
> I have a user who just repeated an experiment for the millionth time but
> using the new formula of Prolong Gold Antifade with Dapi (P36935).  This new
> formula can be kept at room temp, but her former prolong had to be stored in
> the freezer.  As far as we can figure, the new prolong is the only thing
> changed in her protocol.  And unfortunately, instead of imaging beautiful
> FITC-labeled elastin, she saw green elastin and bright green cytosol.  She
> previously had seen in the FITC channel nothing from cells and green fibers
> (elastin) around the cells.
>
> Has anyone tried the new prolong gold?  Because that is the only thing we
> can think of that might be different.  We are going to test by repeating and
> just mounting in PBS.
>
> Thanks,
> Pam
>
> Dr Pamela A. Young | Light and Optical Microscopist
> Australian Centre for Microscopy & Microanalysis
>
> THE UNIVERSITY OF SYDNEY
> Rm 116A, Madsen Building F09 | The University of Sydney | NSW | 2006 |
> Australia
> T [phone] | F [phone]
> E [hidden email]<mailto:[hidden email]> | W
> http://sydney.edu.au/acmm
>
> Incorporating:
> Australian Microscopy & Microanalysis Research Facility (AMMRF) | W
> http://www.ammrf.org.au<http://www.ammrf.org.au/>
> ARC Centre of Excellence for Design in Light Metals | W
> http://www.arclightmetals.org.au<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.
>

--
http://br.linkedin.com/pub/jens-rietdorf/6/4a3/189/
Skype Jens.Rietdorf

[Loralei Dewe-3]

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You need to try FluorGlo by Valley Scientific...made by researchers just
for your specialized applications.

Loralei
 On Jul 17, 2014 7:23 AM, "jens rietdorf" <[hidden email]> wrote:

> *****
> 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.
> *****
>
> Dear Pamela,
>
> I found DAPI concentrations too high by a factor of at least 10.000 in
> the old Prolong, in particular when working with weak other labels
> (why are you using FITC?, is this labeled primary AB? consider to use
> Alexa anti FITC 2ndary to boost the signal), high DAPI conc produces a
> strong background, in particular in the green.
>
>
> Cheers, Jens
>
> On 7/17/14, Pamela Young <[hidden email]> wrote:
> > *****
> > 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.
> > *****
> >
> > Hello Listers,
> >
> > I have a user who just repeated an experiment for the millionth time but
> > using the new formula of Prolong Gold Antifade with Dapi (P36935).  This
> new
> > formula can be kept at room temp, but her former prolong had to be
> stored in
> > the freezer.  As far as we can figure, the new prolong is the only thing
> > changed in her protocol.  And unfortunately, instead of imaging beautiful
> > FITC-labeled elastin, she saw green elastin and bright green cytosol.
>  She
> > previously had seen in the FITC channel nothing from cells and green
> fibers
> > (elastin) around the cells.
> >
> > Has anyone tried the new prolong gold?  Because that is the only thing we
> > can think of that might be different.  We are going to test by repeating
> and
> > just mounting in PBS.
> >
> > Thanks,
> > Pam
> >
> > Dr Pamela A. Young | Light and Optical Microscopist
> > Australian Centre for Microscopy & Microanalysis
> >
> > THE UNIVERSITY OF SYDNEY
> > Rm 116A, Madsen Building F09 | The University of Sydney | NSW | 2006 |
> > Australia
> > T [phone] | F [phone]
> > E [hidden email]<mailto:[hidden email]> | W
> > http://sydney.edu.au/acmm
> >
> > Incorporating:
> > Australian Microscopy & Microanalysis Research Facility (AMMRF) | W
> > http://www.ammrf.org.au<http://www.ammrf.org.au/>
> > ARC Centre of Excellence for Design in Light Metals | W
> > http://www.arclightmetals.org.au<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.
> >
>
>
> --
> http://br.linkedin.com/pub/jens-rietdorf/6/4a3/189/
> Skype Jens.Rietdorf
>