Re: STED slides

Posted by Steffen Dietzel on
URL: http://confocal-microscopy-list.275.s1.nabble.com/STED-slides-tp7588039p7588048.html

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Hi Ali,

what we use for courses and introductions are two color slides:
Alexa594/Atto594/Aberrior Star 580 in one channel and Atto647N/Aberrior
Star 635P/SiR in the other. For example, nuclear pores and actin are a
nice pair of targets. Other than using these dyes (which are less prone
to bleaching under STED conditions than other dyes) this is a normal
immunostaining, so anyone with a little experience, a running cell
culture, and the reagents should be able to do it. EdU replication
labeling also looks cool, but doesn't work together with actin staining.
When I see that Molecular Probes sells slides between 200 and 350 Euro a
piece (not STED slides, I think), it seems that buying the reagents
comes cheaper very quickly.

We usually embed in ProLong Diamond. I am not really happy about it,
because it shrinks the sample along the z-axis when it cures and it
doesn't seem particular intelligent to first shrink and then do
superresolution. But it does provide nice anti-fade properties.

Compared to the 775, we had much less convincing results with the 592. I
hear it is good for YFP, but we didn't try that one ourselves.

Have fun with your new scope!

Steffen


Am 15.03.2018 um 09:45 schrieb Dun, Alison:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
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> *****
>
> Hi
>
> Looking for some training slides for the SP8 3X STED we have just had installed. Have 775 and 592 lasers and want some multicolour prepared slides for training and for media images etc. Anyone know of where I can get some? Leica are low in stock for theirs so need alternative ideas,
>
> Cheers!
> Ali
>
>
> Dr. Alison Dun
> Edinburgh Super Resolution Imaging Consortium (ESRIC)
> 3.29 William Perkin Building
> Heriot-Watt University
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--
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Steffen Dietzel, PD Dr. rer. nat
Ludwig-Maximilians-Universität München
Biomedical Center (BMC)
Head of the Core Facility Bioimaging

Großhaderner Straße 9
D-82152 Planegg-Martinsried
Germany

http://www.bioimaging.bmc.med.uni-muenchen.de