Posted by
George McNamara on
URL: http://confocal-microscopy-list.275.s1.nabble.com/mEos4-photoconversion-tp7583802p7583829.html
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Hi Arvydas,
The sCMOS cameras from (alphabetical order) Andor, Hamamatsu and PCO
have similar pixel size but 5x or 5x as many pixels as your old CCD, so
a lot more area covered. The sCMOS focusing is a lot faster: 100 fps
full frame ... the lower price models are 30 fps (ex. USB3), still
faster than an old CCD.
If you have limited budget, go with an A,H, or P sCMOS that is USB3
(lower price, still 16-bit ... play with contrast) and invest in LED
illuminator, maybe for price consider ThorLabs 4LED
https://www.thorlabs.com/newgrouppage9.cfm?objectgroup_id=3836new filter(s), -- ideally a matched Quad filter set --- and find more
money for a Sutter + Semrock fast emission wheel with threaded filters -
similar cost to slow wheels, but 'time is money".
http://www.semrock.com/how-to-get-the-most-out-of-your-sutter-filter-wheel.aspxthe right LEDs, right Quad set means no moving parts on the excitation
side. Emission filter wheel (especially assuming wide emission bands on
the Quad set) would enable you to choose what fluorophore(s).
If you can find additional money (neighboring labs? friendly dean?)
maybe time to get at least one new objective lens or lenses.
Finally, "instant gratification" deconvolution is just about here --
http://www.microvolution.com/ and
http://www.microvolution.com/technology.php(yes, my image is on their home page - my colleague went "Wow" when he
saw the deconvolved data),
along with NVidia TITAN X card or cards (hey, at least you wouldn't need
to buy a new PC) ... new GPU cards would also let you put HD 4K monitors
on your system. Finally, an data center quality solid state drive for
local file saving (and then transfer to the network to a server for long
term storage and other desktop PC's access).
Do me a favor - for any experiment, do not use overlapping emission
filters like this paper did
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3653935/figure/pone-0063286-g004/the emission filters are about $125 each (a bit more for threaded) -- a
tiny fraction of PLoS One's page charges, to say nothing of the money
spent on salaries for that paper (they also should have multimerized the
mPlum and other dim FPs). If you do like the above paper, see 26010570
for new (one week) paper that cites some of the recent developments in
many color cells ('brainbow 2015'). you may also find of use,
http://works.bepress.com/gmcnamara/65/ and my other writing on
Tattletales and T-Bow.
enjoy,
George
On 6/2/2015 10:12 AM, Arvydas Matiukas 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.
> *****
>
> Hello list,
>
> Please share your experience and advice on CCD camera
> for imaging fixed slides on widefield fluorescent microscope.
> We are looking for a reasonably priced, reliable and sensitive
> camera. High speed is not very important. Also how newest sCMOS
> compare to CCD?
>
> Our 5 year old 1.4MP cooled CCD camera died, and as it
> is discontinued, we prefer to replace it instead of repairing.
> It has been mostly used for imaging fixed brain slices
> labeled by immunofluorescence or fluorescent proteins,
> some labels being quite dim.
>
> Thanks,
> Arvydas
>
> Arvydas Matiukas, Ph.D.
> Director of Confocal&Two-Photon Core
> SUNY Upstate Medical University
> Neuroscience& Physiology Dept
>
>
--
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