*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Re Q2: Does your system auto-adjust exposure time when you 'zoom' or use an
ROI? (sorry, had to ask just in case)  What are the timescales on which
photoconversion of the molecule takes place vs. bleaching?


On Fri, Nov 8, 2013 at 12:38 PM, Feinstein, Timothy <
[log in to unmask]> 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]>
>