Imaging and Flow Cytometry Core CPOS
LSM 800 SOP
E-7
ver. 2.1 (Updated: 11/13/2020)
E.
Locate specimen, focus, find cells / region of interest
1.
You should be on "Locate" tab if the software
has just finished calibration. If not navigate to
"Locate" tab.
2.
Select objective lens with software / on
microscope control panel.
Consult Chapter B to see which objective lens
needs immersion oil.
3.
Push back condenser column.
4.
Mount your sample on sample carrier. Ensure
your specimen holder is level.
5.
[Optional] Click "BF" on software for easier
sample finding and bright-field observation.
6.
Move the area of the slide you wish to observe
above objective lens with X-Y manipulator on air
table.
7.
Focus onto your specimen. Notice "Z-position"
value shown on the microscope control panel
and software, larger number means higher focus
height.
Typical focus height for glass slides ~ 1200
–
1500 µm. Confirm sample placement if out of
range.
8.
Click "B" = Blue; "G" = Green; "R" = Red; "Far-
red" on software for fluorescent observation.
**Note: fluorescent dye will eventually be
bleached under intense excitation light. Find
your region of interest quickly. When you turn
to other tasks turn of ALL illumination by clicking
on "All off" button. **
9.
Click "Acquisition" tab when you arrive at region
of interest.
X
Y
