It is definitely different from phenlyene diamine. It is glycerol based
with a r.i. of 1.4 something. If you are using a water immersion lens that
is made for use with a coverslip, it assumes that the sample is in
glycerol, even if the immersion liquid is water. If your water immersion
lens is made for use without a coverslip, then there are things that you
can add to the buffer to reduce photobleaching, like ascorbic acid.

It is by far the best anti-fade that we have seen.

________________________________________________________________________________


Paul Goodwin
Image Analysis Lab
FHCRC, Seattle, WA

On Thu, 20 Feb 1997, David Knecht wrote:

> Dear Paul- Do you know if Fluoguard is water based or glycerol or what its
> refractive index is?  We want to match a water immersion lens so if it is
> glycerol or the like, it is not of much use.Also, do you really think it is
> different from phenylene diamene or the like?  Dave
>
>  >Margitta-
> >
> >I guess its not clear to me what you mean. The confocal image of FITC is
> >going to show you fluorescence. I don't see how you intend to do
> >densitometry on a fluorescent image. I, instead, you intend to measure the
> >intensity of the brightness of the images, it will be subject to the
> >issues of quantitative fluourescence that have been adressed here lately.
> >In answer to the Antifade, yes, absolutely use it. I would recommend the
> >Fluorgard product from BioRad since it does not appear to quench the FITC
> >signal. I would also look for fields under bright field, then collect the
> >images under fluorescence if at all possible or at least look for cells
> >under PI staining. If you look for bright cells under FITC and collect
> >them with FITC, you will be adding the error of photobleaching caused
> >while looking at the specimens and you will very much bias your data.
> >
> >_______________________________________________________________________________
> >_
> >
> >
> >Paul Goodwin
> >Image Analysis Lab
> >FHCRC, Seattle, WA
> >
> >On Thu, 20 Feb 1997, Margitta Kampman wrote:
> >
> >> I am planning to perform densitometry on tissue sections stained with
> >> FITC-conjugated antibodies. Should I use antifade mountant? Which one
> >> should I chose?
> >> Grateful for any suggestions,
> >> Margitta
> >>
>
>
> Dr. David Knecht
> Department of Molecular and Cell Biology
> University of Connecticut
> U-125
> Storrs, CT 06269
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>