Low-noise microscope CCD camera
- From: Kevin Sunley <umsunle0@xxxxxxxxxxxxxxxxxxxxx>
- Date: Sat, 15 Apr 2006 22:21:04 +0000 (UTC)
Hi everyone,
We're in the process of beginning to look into buying a dedicated
microscope CCD camera for our lab.
This microscope will (initially at least) serve two different and
distinct purposes:
1) a general camera for non-critical photomicrography: including colour
images of stained cells, mostly in brightfield with some phase of DIC
work.
2) quantitative imaging of ratiometric fluorescence of FITC and GFP.
Obviously the first use does not require a particularly advanced camera,
so most of my problems have to do with a camera which can work for both
applications.
AFAIK the quantitative fluorescence work will require as great a bit-
depth as possible (so far I've been looking at 12-bit A/D digital
cameras). Texts I've read which specifically deal with cameras
appropriate with ratiometric imaging seem stress the requirement of
high-speed acquisition, but I believe this is more for measuring very
fast dynamic cellular responses such as changes in Ca2+ levels within
the cell. Our project so far will be looking at using localized FITC
(as a pH sensitive probe) and modified GFP (to make the protein
sensitive to different redox states).
The organelles we hope to measure these conditions in have quite robust
regulation mechanisms to keep their pH and redox states stable so we do
not expect any blindingly fast cellular responses to measure even under
the experimental conditions we plan.
So the question comes down to cooled vs. non-cooled cameras, and colour
vs. monochrome.
Are modern non-cooled colour CCD cameras (such as the Jenoptik ProgRes
C12plus (retail price around $4k)) capable of long enough low-noise
exposures for this type of imaging? Or would a cooled CCD be required
for this type of work? The C12plus's 12-bits per colour channel seems
like they would offer enough data for quantification, but that's
assuming it would not be over shadowed by high noise levels from the
long exposures these applications may require.
The cells we are using are notoriously hard to transfect so the GFP may
be produced in low quantities.
A camera in the same class as the JenOptik would be ideal as it is not
cooled (and therefore less expensive), and could serve both uses we'll
need as it is a colour camera.
A monochrome cooled-CCD would be another option, but colour imaging
would require manual recombining of three different images (red, green
and blue filtered). This would likely be too inconvenient for most
users, even if it would give a higher SNR for the fluorescence work.
A cooled, colour CCD is likely too expensive for our budget (most seem
to be near or over $10k.
The ability to integrate seamlessly into ImageJ would also be
beneficial.
Does anyone have any suggestions or recommendations for a camera
suitable to our uses? Replies with commercial interests are also
welcome.
Thanks in advance,
Kevin
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