Prolong Gold: to solidify or not?

[Ellen T. Arena]

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

I am having conflicting suggestions regarding Prolong Gold anti fade mounting medium from Invitrogen.  After mounting - I have always sealed my coverslips using nail polish to prevent the Prolong from solidifying - but many with whom I work let it solidify.  I am worried that solidifying may change the refractive index over time.  The data sheet from the company says to let solidify (they say 'cure') and the refractive index gradually increases - but then to seal it with nail polish to keep the samples for extended periods of time.

What are people's recommendations for what is best regarding Prolong???  Solidify or not??

Many thanks...

Ellen Arena

------------------------------------------------------
Ellen T. Arena, PhD
Pasteur Foundation Postdoctoral Fellow
Unité de Pathogénie Microbienne Moléculaire
INSERM U786
Institut Pasteur
28 rue du Dr Roux
F - 75724 PARIS Cédex 15
France
Tel: [phone]
Fax: [phone]
[hidden email]
------------------------------------------------------

[lechristophe]

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

I'd say solidify. It is supposed to be a hard-set mountant.

On Mon, Jul 2, 2012 at 12:35 PM, Ellen T. Arena <[hidden email]>wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> I am having conflicting suggestions regarding Prolong Gold anti fade
> mounting medium from Invitrogen.  After mounting - I have always sealed my
> coverslips using nail polish to prevent the Prolong from solidifying - but
> many with whom I work let it solidify.  I am worried that solidifying may
> change the refractive index over time.  The data sheet from the company
> says to let solidify (they say 'cure') and the refractive index gradually
> increases - but then to seal it with nail polish to keep the samples for
> extended periods of time.
>
> What are people's recommendations for what is best regarding Prolong???
>  Solidify or not??
>
> Many thanks...
>
> Ellen Arena
>
> ------------------------------------------------------
> Ellen T. Arena, PhD
> Pasteur Foundation Postdoctoral Fellow
> Unité de Pathogénie Microbienne Moléculaire
> INSERM U786
> Institut Pasteur
> 28 rue du Dr Roux
> F - 75724 PARIS Cédex 15
> France
> Tel: [phone]
> Fax: [phone]
> [hidden email]
> ------------------------------------------------------
>

[Kilgore, Jason-2]

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

** Vendor Reply **

I actually helped develop ProLong Gold.

Though it will work as a mountant and as an antifade before it hardens, you do need to let it harden (or cure) to get the best refractive index.

If you seal the edges prior to curing, it will not cure correctly, since it needs the air interface.  I would wait a day or two before sealing, to make sure curing has completed.

ProLong Gold does not require secondary sealing, unlike the old ProLong, unless you archive the slides.  If you don't secondary seal the edges, then after several months of storage of the slide it will start to have a golden appearance to the edges (where it has oxidized the antifade components).  So if you are archiving your slides long-term, then secondary sealing becomes something that you would need to do.

Cheers,

Jason

Jason A. Kilgore
Technical Application Scientist
Molecular Probes Labeling and Detection Technologies
Cells Systems Division
 
T 1 800 955 6288 then option 4, then option 6,  or  541 335 0353 . F 541 335 0238
29851 Willow Creek Rd . Eugene . OR . 97402-9132 . United States
www.invitrogen.com/technicalsupport

 




-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Christophe Leterrier
Sent: Monday, July 02, 2012 4:05 AM
To: [hidden email]
Subject: Re: Prolong Gold: to solidify or not?

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

I'd say solidify. It is supposed to be a hard-set mountant.

On Mon, Jul 2, 2012 at 12:35 PM, Ellen T. Arena <[hidden email]>wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> I am having conflicting suggestions regarding Prolong Gold anti fade
> mounting medium from Invitrogen.  After mounting - I have always sealed my
> coverslips using nail polish to prevent the Prolong from solidifying - but
> many with whom I work let it solidify.  I am worried that solidifying may
> change the refractive index over time.  The data sheet from the company
> says to let solidify (they say 'cure') and the refractive index gradually
> increases - but then to seal it with nail polish to keep the samples for
> extended periods of time.
>
> What are people's recommendations for what is best regarding Prolong???
>  Solidify or not??
>
> Many thanks...
>
> Ellen Arena
>
> ------------------------------------------------------
> Ellen T. Arena, PhD
> Pasteur Foundation Postdoctoral Fellow
> Unité de Pathogénie Microbienne Moléculaire
> INSERM U786
> Institut Pasteur
> 28 rue du Dr Roux
> F - 75724 PARIS Cédex 15
> France
> Tel: [phone]
> Fax: [phone]
> [hidden email]
> ------------------------------------------------------
>

[obsydion]

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

Although the protocols say that 24 hours is sufficient for solidification, our
microsphere beads appear to swim around even after two full days at room
temperature. Does anybody have a better estimate for complete curing or when
it would be safe to seal with nail polish?

[Kilgore, Jason-2]

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

** vendor reply **

Kelvin, was there any surfactant or detergent, such as Tween-20 or Triton X-100, in with the beads?  This is sometimes added to prevent bead aggregation.  There is some anecdotal evidence that the presence of surfactants can inhibit good curing of ProLong Gold.

Jason

Jason A. Kilgore
Technical Application Scientist
Molecular Probes Labeling and Detection Technologies
Cells Systems Division
 
T 1 800 955 6288 then option 4, then option 6,  or  541 335 0353 . F 541 335 0238
29851 Willow Creek Rd . Eugene . OR . 97402-9132 . United States
www.invitrogen.com/technicalsupport

 




-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Kelvin Poon
Sent: Monday, July 02, 2012 10:36 PM
To: [hidden email]
Subject: Re: Prolong Gold: to solidify or not?

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

Although the protocols say that 24 hours is sufficient for solidification, our
microsphere beads appear to swim around even after two full days at room
temperature. Does anybody have a better estimate for complete curing or when
it would be safe to seal with nail polish?

[David Baddeley]

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


We've found prolong to be quite sensitive to storage, with it not setting properly after it's been in the fridge for a while (a week or two). The solution seems to be to aliquot and freeze on opening the bottle.

You also don't want to have too much excess moisture on the slide before mounting.

Cheers,
David


------------------------------
On Tue, Jul 3, 2012 5:36 PM NZST Kelvin Poon wrote:

>*****
>To join, leave or search the confocal microscopy listserv, go to:
>http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>*****
>
>Although the protocols say that 24 hours is sufficient for solidification, our
>microsphere beads appear to swim around even after two full days at room
>temperature. Does anybody have a better estimate for complete curing or when
>it would be safe to seal with nail polish?