- 131 -
§ Attach waste tubing to pump
§ Insure that waste tubing is going to a waste container
§ Center the fine focus travel if required
5. Setup the software/hardware in the following order:
§ Open Visual Spreadsheet and select the correct objective
§ Insure that the Status Bar color (if displayed) at the bottom of the Main Window matches the
colored stripe on the installed objective
§ Main Window: Setup > Setup and Focus
· Focus on the flow cell using the coarse adjustment Z translator
· Use the Course Adjustment screw on the Z translator to lock the Z translator into place
· Add the sample
and turn on the pump until the sample can be seen in the Setup and Focus
window. Note: Proper sample preparation is critical to having a successful FlowCAM
analysis. It is important to get the proper particle size distribution and sample dilution to
ensure that the best possible data are obtained.
· Use the fine focus adjustment knob to adjust the focus until the sample appears to look
the best possible
· Setup and Focus window: Setup > Camera
· Move the Camera Settings window so that it is not overlaying the Setup and Focus
window (Note: You must click “Apply” for any changes in the Camera window to take
effect).
¨ Adjust the U and V color balance until the displayed camera image is a uniform gray
background (only necessary for a color camera)
¨ Adjust the Gain and Flash Duration (increased gain and flash duration will result in a
higher intensity mean) until you have an intensity mean of 165-185
¨ Close the Camera window
· Determine the concentration, coloration, and size distribution of the sample by looking at
the particles that are displayed in the setup and focus window
§ Close the Setup and Focus window
§ Main Window: Setup > Context
· Context Window: Adjust the context settings via the following tabs:
¨ Capture
¨ Flow Cell