I've built STORM/PALM system and can confirm that it's not t
*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****
Hi David,
I've built STORM/PALM system and can confirm that it's not too difficult. That
said, every system will usually be quite different, depending on the specific
requirements and abilities making a 'one size fits all' instruction manual very
difficult to write. In general it'd be fair to say that some optics/laser design
& alignment experience is desirable when it comes to getting the hardware side
working, and it might be worth getting someone from the local physics department
to help you. Programming experience is also a plus, although something like
'QuickPALM' (as mentioned by John) might give you a leg up (you'd probably still
need to do a bit of tweaking to get everything working on your system). I'm
personally not a huge fan of ImageJ/umanager/ Java in general, so would probably
look for another solution, but this might be personal preference. We use custom,
in house, software which I think is a lot more mature, but which is currently
not particularly well documented.
It also pays to give some thought to what variants of PALM/STORM you want to
support as this has a huge influence on the setup. If your users are looking at
fixed specimens with antibody labelling, something like dSTORM is probably the
easiest to implement (single laser, no complicated switching patterns, can get
away without software control of shutters etc ...).
The two aspects of the hardware that you can't really get off the shelf
components for and are probably going to need to play with are:
- the laser coupling (commercial TIRF couplers typically give too little power
over too large a field of view),
- the focus mechanism (most commercial z-drives show quite a lot of drift - so
you're going to either need some form of active stabilisation, or to redesign
the focus mechanism - we ended up using a PIFoc piezo focusser on a custom metal
bracket & doing away with the microscopes focus mechanism completely).
If you've got any specific questions, you're welcome to get in touch,
cheers,
David
----- Original Message ----
From: David Burk <[log in to unmask]>
To: [log in to unmask]
Sent: Tue, 1 February, 2011 4:05:53 AM
Subject: Building a STORM/FPALM
*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****
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?
*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****
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:
>>
>>> *****
>>> 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
Tel: +39-010.71.781.503
Fax +39-010-72.03.21
Mobile +39-3666719968
www.iit.it
[log in to unmask]
|
