Last updated on Mar 2022
Stage movement is
not smooth/jerks
around
1.
Switch off the stage controller
2.
Manually manipulate the stage to each of the 4 corners
3.
Spray compressed air to dislodge any dust particles that may be
causing issues.
4.
Switch on the stage controller and restart the Fluoview
software
5.
Select “Yes” when you see the popup (shown below) asking if
you want to execute the cleaning process for the stage unit.
No signal during
confocal scanning
1.
Use the transmitted detector to check. If the specimen is thick
and non-transparent, the specimen should be removed to first
check for laser pass through to transmitted detector. For easier
verification, standard specimen slides can be used as
confirmation check.
2.
Check to see if scanning spot can be spotted on the specimen.
3.
If there is no scanning spot:
I.
Check that main laser key is switched on and the
appropriate laser is switched on.
II.
Check the laser rack to check the laser status light.
Blinking light on the diode laser (which indicates laser
starting up) has turned to a solid white or coloured light
(sometimes red light might be indicative of error).
III.
Restart the main controller right at the bottom if none of
the steps taken work, especially if the machine has been
left on by the previous user.
Signal image not
visible/too dim
even at high laser
power
Check
: If fluorescence signal is visible on wide-field eyepiece.
Check
: PMT settings if fluorescence can be seen on wide-field
Ideal base settings are
•
HV
: 350-500V for HSD, 600-800V for SD.
You can set the HV
higher for quick sample spotting, then when you find the sample
you can tune the HV lower and increase laser power.
•
Gain:
1x.
Decreasing Gain reduces brightness and increasing
Gain doesn’t increase the sensor’s sensitivity anyway.
•
Offset:
0%.
Adjust it only after you find your sample
.
•
Pinhole:
Auto.
When set to manual mode it may default to the
smallest size which will make the signal very dark.
Control panel
switches off when
Live View is
activated
This is a feature by Olympus to allow a fully dark room during imaging
that can be switched on/off when needed.
To check status, go to Configuration > Preference tab > Microscope Link
and check
Turn the TPC backlight off during scanning
and
Turn the LED
indicator off during scanning
. You can set them to your preferences.
