Posted by
Zdenek Svindrych-2 on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Elyra-7-system-for-bioluminescence-tp7591945p7591949.html
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Hi Michal,
you may want to use much longer exposure times (30 seconds for CMOS
cameras, hours for CCDs) depending how weak your luminescence is. With
EMCCDs, since the readout noise is negligible, you should get the same
results by summing shorter exposure images, like you suggested. You may
also bin the pixels for more SNR.
For optimization and troubleshooting you can measure the luminescence from
your cells with a suitable plate reader, they are much more sensitive.
I haven't tried this with the Zeiss Axio stand, but one issue I found with
the Nikon Ti-E and TE2000 is the stray light coming from internal
components (probably the encoders). I had to power off the microscope body
completely to get low background (I think Ti2-E can power off the encoders
only, if needed).
Also, dedicated systems may have much better light throughput, especially
when you trade off resolution for collection efficiency (just compare the
back focal plane aperture size of your microscope objective lens with let's
say Nikkor 50 mm F/# 1.2.
Good luck!
Best, zdenek
On Mon, Mar 8, 2021 at 8:50 AM Michał Majkowski <
[hidden email]> wrote:
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>
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>
> Dear List Members,
> I am just thinking about the use of our Elyra 7 system to visualise
> bioluminescence.
> Have any of you ever tried something similar? In systems dedicated for
> bioluminescence signal is detected with the same or similar camera (Andor
> iXon Ultra 897) though light path is less complex.
> My idea is to take time series with longest possible exposure time (1 sec),
> convert T->Z and then sum up all slices. Does it have any sense?
> Thank you in advance
> Michal Majkowski
>
> Imaging Specialist
> University of Wroclaw
> Biotechnology Faculty
> Joliot-Curie 14a
> 50-383 Wroclaw POLAND
>
--
--
Zdenek Svindrych, Ph.D.
Research Scientist - Microscopy Imaging Specialist
Department of Biochemistry and Cell Biology
Geisel School of Medicine at Dartmouth