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Hi Valeria,
I recommend a 4 megapixel or 5.5 megapixel scientific CMOS camera, for
examples from Hamamatsu (FLASH4.0), PCO or Andor. these have the same
pixel size (~6.8x6.8um) but 3x more pixels than a typical CCD camera (ex
ORCA-ER) so 3x bigger field of view, and sCMOS have much faster focusing.
I've been using Bruce & Butte GPU deconvolution for (almost) instant
gratification deconvolution
http://www.opticsinfobase.org/oe/fulltext.cfm?uri=oe-21-4-4766&id=249375
See
http://works.bepress.com/gmcnamara/37/
and other FISH data posted at http://works.bepress.com/gmcnamara
for more examples. I am using the late 2013 NVidia TITAN card. The
newest TITAN X should be somewhat faster.
The free B&B Deconvolution is no longer available - see
http://www.microvolution.com/ for the commercial version (disclosure: I
was involved in acquiring the image on their home page).
See http://stellarisfish.smugmug.com/ for the single RNA
molecule FISH vendor galleries (Biosearch Tech). If the probe set you
want is not part of BTI's catalogue line, takes about a week to make and
ship 'custom' set. With my microscope (Lumencor SOLA in single LED mode,
Leica DMI6000, 6 filter cubes) I recently found Quasar 705 is somewhat
brighter than Quasar 670 for the same probe set. Quasar 570 and CAL
Fluor Red 610 work fine. I avoid ordering probe sets in the green since
the pancreatic cancer cells I FISH have high green fluorescent
autofluorescence.
Enjoy,
George
On 4/7/2015 8:27 AM, Valeria Berno wrote:
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Dear all,
>
> just a quick survey....if I need to use a widefield microscope dedicated to acquire DNA e RNA FISH slides on cell with 100X obj (all of these experiments are either with home-made probes (not very strong signal) and small cell (lymphocytes))......
>
> which source light (LED, Xenon..), but mainly which type of camera would you suggest? or even better which are the camera characteristics I cannot underrate?
>
> Thanks in advance for all your always useful replies.
>
> Valeria
>
>
> Valeria Berno, PhD
> Imaging Facility
> ______
>
> Istituto Nazionale di Genetica Molecolare
> Istutito Nazionale Genetica Molecolare "Romeo ed Enrica Invernizzi"
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--
George McNamara, Ph.D.
Single Cells Analyst
L.J.N. Cooper Lab
University of Texas M.D. Anderson Cancer Center
Houston, TX 77054
Tattletales http://works.bepress.com/gmcnamara/42
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