Posted by
Stanislav Vitha-2 on
URL: http://confocal-microscopy-list.275.s1.nabble.com/laser-alignment-issue-FluoView1000-tp7584981p7585282.html
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Sometimes, the motorized slit in the spectral detector unit gets stuck, so if
it is stuck in a narrow bandwidth setting, you would be getting very little
signal. But it is unlikely that this would happen on both detectors the same
time.
You could try changing the bandwidth from the maximum 100 nm down 10
nm when imaging a fluorescent slide or a solution of a fluorescent dye. If
the slit adjustment works, the signal will go down.
Another possibility is that the spectral calibration is off, but again not likely
to happen for both detector units, unless somebody played with the
settings, logged as Maker (administrator). One easy way to test is to image
a surface in reflected mode, with minimum spectral bandwidth (2nm). For
instance, use 488 nm laser, set the detector to 488-490nm detection. In
live mode, start moving the detector band in 1 nm steps up or down and
watch the signal intensity. Once you get out of the laser wavelength range,
the signal should drop very fast.
Stan
Dr. Stanislav Vitha
Texas A&M University
Microscopy and Imaging Center