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You need to set the detector range to match the dimmest and brightest signal
from the specimen. Setting the detector incorrectly results in the loss of
information from the specimen.
3. Set
Ampl. Offset
– setting the ‘min’ signal
This should be set next, but may need re-setting later. While acquiring with the
Fast
scan
button, adjust the
Amp. Offset
so that only a few of the pixels in the background
of the image are blue. If you have a lot of blue colour in the background, move the slider
to the right.
4. Set
Detector Gain and Excitation
power
– setting the ‘max’ signal
The ‘max’ signal is set by adjusting the
detector gain
and
laser
transmission
simultaneously. You
need to empirically work out the best
laser power settings – low laser power
causes less bleaching but requires the
detector gain to be set high; which
introduces noise.
Whist acquiring with the
Fast scan
button, adjust increase the
Detector Gain
so that
you get a bright image, but not too many red (saturated) pixels and not too much noise.
You may need to adjust the
Ampl. Offset
if you increase the
Detector Gain
a lot.
Around 600 is a good start. This should be set in conjunction with the excitation
intensity.
5. Set Ampl. Gain
Leave as 1 unless you have a very dim signal and nothing else works. This will amplify
noise as well as signal.
