Start-up
1 . . . . . . . . . Turn ON Master Switch
2 . . . . . . . . . . . Log on to Computer
3 . . . . . . . . . . . . . Load Software
Optional . . . . .Turn ON X-Cite Hg Lamp
Align Optical Components
1 . . . . . . . . . . .Koehler Illumination
Calibrate System
1 . . . . . . . . . . . . . Grid Tune Lens
2 . . . . . . Parcentric Parfocal Calibration
Run Experiment
1 . . . . . . . . . . . . . .Set up Subject
2 . . . . . . . . . . Set scope to Low Mag
3 . . . . . . . . Trace Region(s) of interest
4 . . . . . . . . . .Set scope to High Mag
5 . . . . . . . Define Counting Frame Size
6 . . . . . . . . . Define SRS Grid Layout
7 . . . . . . . . . Define Disector Options
8 . . . . . . . . Save Sampling Paramters
9 . . . . . . . Select Markers for Counting
10 . . . . . . . . . . . . .Count Objects
11 . . . . . . . . .View Sampling Results
Save Files
. . . . . . . . . . . . .
1
. . . . .Save to local machine-(often)
2 . . . . . . . Copy Files Server (Zeus)
1. Koehler align optic train.
2. Deflect light to camera with push
pull rod located above binocular vertical
control knob.
3. Start StereoInvestigator. Select Imag-
ing > Live Image (Ctrl-L)
4. Click with mouse inside image frame
to set reference point and to improve
performance of mouse when hovering
over the display area.
5. Select Tools > Video > Camera Set-
tings... to bring up Camera Settings win-
dow to help with setting exposure Gain
and other camera specific options.
6. Adjust Exposure and white balance
using More... button in the Camera Set-
tings window, another window will pop
up with the basic Olympus PictureFrame
software interface “Auto exposure” and
“AWB” auto white balance are the two
most important functions. Ensure to
select an area for white balance in the
Olympus PictureFrame “live window.”
With StereoInvestigator
Acquire Images
7. Create a flatfield (optional)
Find a clear area for bright field, or even-
ly fluorescent field for epi-fluorescence;
fluorescence dye slides are available on
request.
Capture image: Imaging > Acquire
Image. Set the captured image as a
background image:
Flatfield for Brightfield: Imaging >
Set to Brightfield Background Image
or
Flatfield for Fluorescent: Imaging >
Set to Fluorescent Background image
Tell software to subtract background
image during acquisition: Imaging > En-
able Background Correction.
8. Acquire images: Imaging > Acquire
Image. The acquired image will replace
the live image remember to select Imag-
ing > Live Image (Ctrl+L) to return to live
viewing of sample.
9. Save Images select File > Image Save
or Image Save As to save captured im-
ages.
