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Hi Tim,

I disagree with your statement "lacking in sensitivity". The quantum 
efficiency of your ORCA-2.8 is better than the ORCA4.0 at less than 500 
nm, and better than "Gen 1" sCMOS at less than ~570 nm. See Spectral 
Response graph at
http://www.hamamatsu.com/jp/en/community/life_science_camera/product/search/C11440-22CU/index.html
and SNR curve next to it.

The Huang et al method for full calibration of every pixel should apply 
to all sCMOS cameras and deal with the pixel specific gains (which is 
not really noise).

Video-rate nanoscopy using sCMOS camera-specific single-molecule 
localization algorithms. <http://www.ncbi.nlm.nih.gov/pubmed/23708387>

Huang F, Hartwich TM, Rivera-Molina FE, Lin Y, Duim WC, Long JJ, Uchil 
PD, Myers JR, Baird MA, Mothes W, Davidson MW, Toomre D, *Bewersdorf J*.

Nat Methods . 2013 Jul;10(7):653-8. doi: 10.1038/nmeth.2488.  PMID: 23708387


You can achieve the optical equivalent of binning by not using the 
highest magnification objective lens.

There are specific applications where back illuminated EMCCD vs sCMOS, 
one will win. For example, UAIM is EMCCD specific:

Ultrahigh accuracy imaging modality for super-localization microscopy. 
<http://www.ncbi.nlm.nih.gov/pubmed/23455923>

Chao J, Ram S, Ward ES, Ober RJ.

Nat Methods. 2013 Apr;10(4):335-8. doi: 10.1038/nmeth.2396.  PMID: 23455923


I recommend you put the FLASH2.8 on the microscope that makes the most 
sense and if your customer base needs another camera system, select 
whatever camera type, and /manufacturer/model fits that need.


mEos4 - still unpublished, except for a poster I saw in Loren Looger's 
lab (and maybe some online abstracts that Google has not found). Try 
contacting Maria at 
https://www.linkedin.com/pub/maria-gabriela-paez-segala/61/978/b55 or 
Loren at HHMI Janelia Farm for more details.


Sincerely,


George


On 11/8/2013 1:38 PM, Feinstein, Timothy wrote:
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Hello all,
>
> I have two questions, bundled into one email to save space.  Any help on or off-list is much appreciated.
>
> 1. Not precisely confocal, but related enough.  As the new core manager I am bringing quantitative widefield, TIRF and in particular FRET to what was previously an exclusive confocal core.  This will call for a good CCD but at the moment we have a Hamamatsu Orca 2.8 SCMOS, which is nice for speed but lacking in sensitivity and un-binnable.  As I recall EM-CCDs have the best sensitivity but a problem with non-Gaussian noise.  Is this still true?  If you had to choose one model for the best combination of sensitivity and quantitative response, what would you choose?  Thanks in advance.
>
> 2. While helping several users with photoconversion of mEOS2, we have noticed an interesting relationship between laser intensity and ROI scan speed.  In our hands it seems like turning the 405nm laser up mostly increases bleaching, whereas effective photoconversion depends on finding a long enough pixel dwell time.  Maybe I am just showing my lack of experience with photoconversion, but it seemed like a counterintuitive result.
>
> Also, I just heard about mEOS4 on this list for the first time.  Does its increased stability mean anything for live imaging, or does it mostly help with post-fixative applications?
>
> Thanks and all the best,
>
>
> TF
>
>
> Timothy Feinstein, Ph.D. | Confocal Manager, Van Andel Research Institute
> 333 Bostwick Ave., N.E., Grand Rapids, Michigan 49503
> Phone: 616-234-5819 | Email: [log in to unmask]<mailto:[log in to unmask]>
>
>    


-- 



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/26/