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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]>