http://confocal-microscopy-list.275.s1.nabble.com/what-fluorescent-proteins-are-working-best-for-precision-localization-microscopy-nanoscopy-in-your-a-tp7580157p7580177.html
Thanks, Doug. For some reason I thought STORM had a more limited dye selection.
Dept. of Pharmacology & Chemical Biology
> *****
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>
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy> *****
>
> This is STORM. It doesn't work with every dye, but many. See here:
>
> Evaluation of fluorophores for optimal performance in localization-based
> super-resolution
> imaging<
http://www.nature.com/nmeth/journal/v8/n12/full/nmeth.1768.html>
>
> - Graham T Dempsey<
http://www.nature.com/nmeth/journal/v8/n12/fig_tab/nmeth.1768_ft.html#auth-1>
> ,
> - Joshua C Vaughan<
http://www.nature.com/nmeth/journal/v8/n12/fig_tab/nmeth.1768_ft.html#auth-2>
> ,
> - Kok Hao Chen<
http://www.nature.com/nmeth/journal/v8/n12/fig_tab/nmeth.1768_ft.html#auth-3>
> ,
> - Mark Bates<
http://www.nature.com/nmeth/journal/v8/n12/fig_tab/nmeth.1768_ft.html#auth-4>
> - & Xiaowei Zhuang<
http://www.nature.com/nmeth/journal/v8/n12/fig_tab/nmeth.1768_ft.html#auth-5>
>
> Nature Methods 8,1027–1036(2011)doi:10.1038/nmeth.1768
>
> -Doug
>
>
>
> On Thu, Apr 11, 2013 at 10:20 AM, Tim Feinstein <
[hidden email]> wrote:
>
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
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>>
>> I wonder whether anyone has experience with the inducible dark states
>> approach reported by Christian Soeller in New Zealand. In theory one could
>> do super-resolution TIRF with any fluorophore you want.
>>
>>
http://www.sciencedirect.com/science/article/pii/S0006349508000763>>
>> Markus Sauer followed up with a protocol:
>>
>>
http://www.nature.com/nprot/journal/v6/n7/abs/nprot.2011.336.html>>
>> Has anyone tried this?
>>
>> cheers,
>>
>>
>> TF
>>
>> Timothy Feinstein, PhD
>> Visiting Research Associate
>> Laboratory for GPCR Biology
>> Dept. of Pharmacology & Chemical Biology
>> University of Pittsburgh, School of Medicine
>> BST W1301, 200 Lothrop St.
>> Pittsburgh, PA 15261
>>
>> On Apr 11, 2013, at 8:35 AM, Tom Blanpied wrote:
>>
>>> *****
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>>>
>>> Hi,
>>>
>>> We use mEos whenever possible. mEos2 seemed great to us, as we never saw
>>> any effects of the small dimerization tendency. mEos3 does look to be at
>> least
>>> as good, and it does seem a bit brighter; it is our go-to probe now. We
>> avoid
>>> the bulk of the tdEos, but did think it was a good molecule.
>>>
>>> PA-mCherry we thought was pretty terrific at first, but we no longer use
>> it
>>> unless we really need the separate GFP channel for something. It is
>>> considerably dimmer in our hands than mEos (photons/frame and number of
>>> frames till bleaching), and also seems to have a tendency to produce a
>> smaller
>>> population of photoactivated molecules. We've not tried to determine
>> whether
>>> this was due to expression level or folding or a non-activatable
>> fraction, but it
>>> does not seem to be due simply to the diminished ability to localize dim
>> or
>>> quickly bleached molecules.
>>>
>>> PA-GFP and Dronpa we did a little bit with, but in general we have so
>> much
>>> autofluorescent green stuff in our relatively old cultured neurons that
>> anything
>>> in that spectral range will be unsatisfying. We finally managed to
>> obtain some
>>> PS-CFP2, but haven't yet measured much about it; we're hopeful it will
>> be the
>>> best of that range.
>>>
>>> I'll be curious to hear comparisons of dendra2-type molecules.
>>>
>>> Tom
>>