Imaging and Flow Cytometry Core CPOS
LSM 800 SOP
F-8
ver. 2.1 (Updated: 11/13/2020)
F.
Confocal imaging set up
Prerequisites: Region of interest is centred in field of view and have clicked 'Acquisition' tab.
1.
Select one of the pre-set made by Imaging and
Flow Cytometry Core.
2.
Go to dropdown menu in "experiment settings"
area. Select settings with prefix "0IFCore_"
3.
Please note "Blue" "Green" "Red" are general
broadband wavelength setting you will need to
fine tune the detection range in Imaging set up
window. (Step F8)
4.
Go to "Imaging setup" window
Decide on switch track every:
“Line”
= change color every line of pixel
“Frame”
= Each color per frame sequential.
“Frame Fast”
= lock up pinhole and wavelength
splitter but still change color per frame.
“Full Z
-
stack”
=
(only available when ‘z
-
stack’ is
activated. Will change track after finishing whole
z-stack.
5.
Select any one track for set up.
6.
Table's rows indicate detector choice.
Row 1 = PMT1
Row 2 = Airyscan detector
(when idle will work as confocal)
Row 3 = PMT2
7.
1st column indicates activation.
8.
2nd column indicates dye reference and name
(click on
for dye selection window).
9.
After setting up dye selection, spectra window
will be updated. Use slider to have detector
detect most of the emission wavelength range.
10.
3rd column indicates pseudo-colour choice.
Click
to change.
11.
Last column indicates detection range.
12.
If a bright-field image is required, check the
'ESID' option in ‘Imaging Setup’ window.
13.
Go to 'Channels' window. While you make
changes please observe histogram in 'Display'
tab under preview area.
Good
14.
For detailed description of 'Display' tab please
read ZEN Blue 2.3 SOP on Imaging and Flow
Cytometry Core website-> Protocols -> Imaging
F7
F8
F10
F11
F12
F6
F5
F9
F4
