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Since the topic of home built super-resolution systems has been brought up I was wondering if anyone had a very simple "Super Res for Dummies" type manual for building such a system. While I feel quite capable of following directions and assembling components together I have a certain degree of trepidation when it comes to figuring out how to drive all the components software-wise. I will admit I lack programming experience and limit myself to making ImageJ macros (poorly) or Cell Profiler pipelines - not writing code in Matlab to open shutters and such. I am aware that many articles describe their setups and give overview diagrams but, from a true schematic standpoint, they lack sufficient detail for someone as rigidly OCD as myself. Perhaps there is a class or course offered somewhere that covers this "roll your own" approach and I have yet to convince Google to divulge that information.
Along the same lines, I am very interested in trying to construct an optical projection tomography system for our facility and, again, while I know many labs have built their own systems and published details of them, some critical details elude me. Have any listers built their own OPT rig and, if so, could you provide a detailed component list as well as assembly instructions? Personally, I would be more than willing to come visit a lab that has implemented their own solution to the FPALM/STORM and/or OPT method if they wouldn't mind spending a little time answering what they most likely would think are silly questions (and would allow me to take pictures of their system).
David
David H. Burk, PhD
Cell Biology and Bioimaging Core
Pennington Biomedical Research Center
Baton Rouge, LA 70808
-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]] On Behalf Of Alberto Diaspro
Sent: Saturday, January 29, 2011 10:47 AM
To: [log in to unmask]
Subject: Re: Who has purchased a fluorescence nanoscope?
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At the Italian Institute of Technology, we got a Leica STED-CW and home built an FPALM on an inverted Nikon thanks to software sharing from Sam Hess Lab, University of Maine. We home built a WLL and a CW STED controlled by MPI Nanobiophotonics software. We are currently interested n the Nikon N-STORM.
All the best
Alby
Il giorno 29/gen/2011, alle ore 17.19, Christophe Leterrier ha scritto:
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> The Bordeaux Imaging Center in Bordeaux, France has a STED and a
> TIRF/PALM http://www.bic.u-bordeaux2.fr/
>
> PALM/STORM setups are becoming more common (I'm aware of 2 setups
> already running in Marseille), because (at least this is what the
> people who did it told me) it is quite easy to add to an existing TIRF
> setup (provided you find software for the detection and localisation
> of individual fluorophores, but there is now an available ImageJ plugin for that).
>
>
> --
> Christophe Leterrier
> Postdoc
> INSERM UMR641 // Ionic channels Lab
> IFR Jean Roche, Mediterranée University Marseille, France
> [log in to unmask]
>
>
>
>
> On Sat, Jan 29, 2011 at 16:52, G. Esteban Fernandez <
> [log in to unmask]> wrote:
>
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> *****
>>
>> UCLA has a STED
>>
>> http://clms.cnsi.ucla.edu/cnsi/clms/equipment-list?search_lab=6791
>> <http://clms.cnsi.ucla.edu/cnsi/clms/equipment-list?search_lab=6791>
>> http://www.cnsi.ucla.edu/staticpages/core-facilities#alms
>>
>>
>> <http://www.cnsi.ucla.edu/staticpages/core-facilities#alms>
>>
>> On Sat, Jan 29, 2011 at 7:32 AM, George McNamara
>> <[log in to unmask]>wrote:
>>
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>>> To join, leave or search the confocal microscopy listserv, go to:
>>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>>> *****
>>>
>>> Dear Confocal listserv,
>>>
>>> Who has purchased a fluorescence nanoscope? What can you tell me
>>> about
>> your
>>> experiences - either here or privately?
>>>
>>> I see 11 OMX labs listed at http://www.api.com/omx-labs.asp (may be
>>> out
>> of
>>> date or some customers may be shy).
>>>
>>> I am aware of STED systems in the USA at Yale Univ, NIH and I was
>>> told
>> San
>>> Diego (have not found where), and possibly UC Denver. Paul French
>> apparently
>>> did his own upgrade of a Leica SP2 (
>>> http://www.focusonmicroscopy.org/2008/PDF/070_Auksorius.pdf and a
>>> Leica Scientific Forum video ... see also related work at
>>> ftp://ftp.espci.fr/incoming/Gilles/INSERM-STED-x6.pdf ).
>>>
>>> I am also aware of one 4pi microscope in the USA.
>>>
>>> So, who has N-SIM, N-STORM, SR-SIM, PALm, SR-200, and what can you
>>> tell
>> me
>>> about your experiences with your (purchased) nanoscope(s)?
>>>
>>> Sincerely,
>>>
>>> George
>>> p.s. Please no need to clutter up the listserv with other people's
>>> nanoscope references - I know how to use PubMed. For that matter,
>>> I've replicated the results of the following two papers on my confocal's:
>>>
>>> Subdiffraction fluorescence imaging of biomolecular structure and
>>> distributions with quantum dots. </pubmed/20600360>
>>>
>>> Heidbreder M, Endesfelder U, van de Linde S, Hennig S, Widera D,
>>> Kaltschmidt B, Kaltschmidt C, Heilemann M.
>>>
>>> Biochim Biophys Acta. 2010 Oct;1803(10):1224-9. Epub 2010 Jun
>>> 23.PMID: 20600360
>>>
>>>
>>> Quantum dot triexciton imaging with three-dimensional subdiffraction
>>> resolution. </pubmed/19453186>
>>>
>>> Hennig S, van de Linde S, Heilemann M, Sauer M.
>>>
>>> Nano Lett. 2009 Jun;9(6):2466-70.PMID: 19453186
>>>
>>> I have used <1 Airy unit pinhole to improve triexciton resolution
>> further.
>>> I have not tried doing 3D deconvolution on the data.
>>>
>>>
>>> --
>>>
>>>
>>> George McNamara, PhD
>>> Analytical Imaging Core Facility
>>> University of Miami
>>>
>>
ISTITUTO ITALIANO
DI TECNOLOGIA
Prof. Alberto Diaspro
Scientific Head
Nanophysics
Via Morego, 30 16163 Genova
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