Posted by
Sudipta Maiti on
URL: http://confocal-microscopy-list.275.s1.nabble.com/2P-standard-tp5923717p5928658.html
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We use fluorescent solutions. We keep a stock in ethanol in the
refrigerator, and dilute it to water before use. We measure the absorbance
of the final solution in a spectrophotometer evertytime. A droplet on the
cover slip is enough to get counts.
Sudipta
On Fri, 14 Jan 2011 18:05:02 -0700, Craig Brideau wrote
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>
> I use the Chroma slides and fluorescein solution at known concentrations.
> Use a water dipping lens with the solution and image away from
> either the bottom of the dish or the surface of the water. You want
> to be about middle depth. It won't bleach and as long as you weight
> it out carefully before putting it into solution you can repeatably
> create it.
>
> Craig
>
> On Fri, Jan 14, 2011 at 5:39 PM, Mark Cannell
<
[hidden email]>wrote:
>
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> >
> > How about Fluorescent plastic slides -I got some free from Chroma.
> >
> > Cheers
> >
> >
> >
> > On 15/01/2011, at 12:34 PM, Boswell, Carl A - (cboswell) wrote:
> >
> > Hi all,
> >> I'm looking for some trustworthy, permanent standard(s) to use in a MP
> >> system to monitor the condition of detectors and the system in general.
> >> The goal is a fluorescent source that is immutable. This could done
either
> >> by having unrestricted replacement of bleached label with unbleached
label
> >> (i.e. a solution), or a uniform and very large solid, so that it is
unlikely
> >> that any one tiny volume will be imaged repeatedly. We've tried
several
> >> iterations of an aqueous solution of label, but sealing the prep does
not
> >> prevent eventual evaporation. Plus, there seems to be some
modification of
> >> FITC, for example, in solution over time (a month) regardless of
exposure
> >> to light. I'm not sure I can trust the consistency of colored plastic
> >> slides, since they are not manufactured with this role in mind. One
> >> possibility is uranyl glass slides, but Corning says they don't make
these
> >> any more. Does anyone know a source? We're also considering quantum
dots
> >> in something like immersion oil , in a sealed chamber but I don't have
any
> >> experience with these labels in a hydrophobic environment.
> >>
> >> Thanks for your insights.
> >> C
> >>
> >>
> >> Carl A. Boswell, Ph.D.
> >> Molecular and Cellular Biology
> >> Univ. of Arizona
> >> 520-954-7053
> >> FAX 520-621-3709
> >>
> >>
> >>
Dr. Sudipta Maiti
Associate Professor
Dept. of Chemical Sciences
Tata Institute of Fundamental Research
Homi Bhabha Raod, Colaba, Mumbai 400005
Ph. 91-22-2278-2716 / 2539
Fax: 91-22-2280-4610
alternate e-mail:
[hidden email]
url: biophotonics.wetpaint.com