Symptom
Possible Causes
Recomended Solutions
No Images
Camera is not running
Click Run/Setup.
If the camera is already
running
Click Stop to stop the camera, and then click Run/Setup.
Imaging is paused
Click Resume.
Displayed region is incor-
rect
In the cell view area, select the all population.
Insufficient illumination
Turn the appropriate lasers on. Set the 785 SSC laser to 40 mW. Set the
laser powers to maximum and decrease them to prevent pixel saturation.
Make sure the brightfield lamp is turned on and click Set Intensity.
Core stream is outside
the objective’s field of
view
Manually find the core stream. In the focus and centering section, move
core track left or right to find the core.
Computer resources are
being over used
Close all third party software.
Imaging and
acquisition rate
is erratic, or
appears frozen
Sample concentration is
low
Make sure the sample concentration is between 107 and 108 cells/mL.
Lower concentrations can be used but this will decrease the cells/second.
Region being viewed has
few or no cells
In the cell view area, select the all population, or readjust regions to include
more cells.
Insufficient illumination
Turn the appropriate lasers on. Set the 785 SSC laser to 40 mW. Set the
laser powers to maximum and decrease them to prevent pixel saturation.
Make sure the brightfield lamp is turned on and click Set Intensity.
The sample is too con-
centrated
The process of object detection can safely handle up to 4000 objects per
second. The maximum sample concentration is 4-5x10^8 cells per mL, with
the recommended concentration 1-10 x10^7 cells per mL. To decrease the
event rate, dilute the sample.
The sample has an
excessive amount of
debris
Use a region to eliminate the debris from the data file, or prepare a fresh
sample.
Computer resources are
being overused
Exit all third party programs.
Objects appear
streaked
Camera is not tracking
the cell velocity accur-
ately
Verify brightfield is working normally and rerun ASSIST using calibration
For more information, see Unstable fluidics (Air or clog in system)
Objects are not
centered in the
channel
Lateral deviation of the
core stream due to air or
clog in the system
Run the purge bubbles script from the instrument drop-down menu.
more information, see Unstable fluidics (Air or clog in system)
Autofocus and centering
is not tracking properly
In the Focus and Centering section, adjust focus and centering left or right,
until the images are centered and in optimal focus.
For Research Use Only. Not for use in diagnostic procedures.
48
Amnis
®
FlowSight
®
Imaging Flow Cytometer User Manual
