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Hi Michal,

I don't know the MRC 600 at all, but if I saw this symptom on my
microscope I'd suspect an off-center pinhole (in addition to the other
suggestions already offered by others).

If you don't feel like trying to center the pinhole yourself, you can
test this idea anyway-- try opening the pinhole little by little and
re-scanning. Intensity always increases when you open the pinhole, but
if you see a *sudden* jump in intensity, your pinhole may be off-center
and missing the in-focus peak of light.

Hope this helps --
Theresa

>> The problem consist in getting not-so-well-focused images on screen,
>> while the cell viewed by eye appears super sharp.  We tested it on
>> phalloidin-labelled cells: we see sharp stress fibres and striations
>> in
>> cardiomyocytes and the same stress fibres appear slightly blurred on
>> the
>> monitor screen and striations are hard to make up.
>>
...
>>      Dr. Michal Opas
>>      Department of Laboratory Medicine and Pathobiology
>>      University of Toronto

==================================
Theresa C. Swayne                         [log in to unmask]
Optical Microscopy Facility             (212) 305-2281
Columbia University, New York, NY

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