40 – LSM 880 Training Guide
Using AiryScan
FAST
In contrast to typical confocal imaging where you fill the dynamic range for optimal contrast; with AiryScan
FAST
we want to set
our sensitivity settings to fill 50% of the total available dynamic range.
1.
From the Acquisition toolbar, expand the blue ‘Channels’ dialog.
2.
Set laser power to a reasonable value for your fluorophore.
Note: The laser power in the FAST mode is reported differently compared to
the base LSM. The first value is the percentage laser power for the FAST
mode and is equivalent to the second value in the LSM mode. For example,
2% of 561nm in FAST mode is equivalent to 0.25% in LSM mode.
3.
Start ‘Live’ scan.
4.
Set ‘Display’ histogram below image to display 50% of dynamic range by
setting ‘White’ point to 128 (8 bit).
5.
Turn on range indicator in Dimensions tab.
6.
Adjust ‘Master Gain’ until brightest areas of the image are just below
saturation.
Note: For further explanation on the use of the range
indicator, please see page 17.
7.
Repeat steps 2-6 for all tracks.
8.
Align AiryScan
FAST
(if not done previously for this sample) –
see page 33 for procedure.
