Posted by
Colin Coates on
URL: http://confocal-microscopy-list.275.s1.nabble.com/flash-4-0v2-for-single-molecule-imaging-tp7582355p7582360.html
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**commercial response**
Hi Jeff,
Bottom line, if you happen to have access to an EMCCD that's generally
what I'd recommend for dynamic single molecule TIRF. After a few years of
our single molecule imaging users comparing both our EMCCD and sCMOS
cameras, the feedback has been pretty convincing. Its generalising, but the
light levels involved in single molecule tend to be below the practical cross
over point in the SNR vs photon flux curve, whereby an EMCCD has the
advantage. Localisation super-resolution microscopy can be the exception,
since the approach requires you to drive up photon levels to be successful.
Having said that, if you are trying to adapt an sCMOS to dynamic single
molecule, one option is to at least bring the effective pixel size closer to
what you are used to with a larger pixel EMCCD. The best way is to use
between 1.5x to 2x demagnification before the camera (depends on the
objective as to what you can get away with and still oversample).
Alternatively, 2x2 binning of the 6.5 um pixel (yielding a 13um binned
pixel) can also be effective. The photons per pixel will quadruple, but be
aware the read noise will also double - however, there's still a net gain in
SNR.
Best,
Colin
Dr Colin Coates
Product Manager - Imaging
Andor Technology