3. Observe the plots for the scatter channel and fluorescent channels. There should be two distinct peaks for
each scatter channel and three distinct peaks for each fluorescent channel. The coefficient of variation (CV)
for bright peaks should be narrow. If either the CVs are very wide on a detector or no signal is observed, the
fluidics need to be primed as described in the following procedure.
To prime the fluidics:
1. Move the tube support arm to the side.
2. Remove the tube from the SIP or leave the HTSconnected to the SIT.
3. Press the PRIME fluid control button to force the fluid out of the flow cell and into the waste container.
Once drained, the flow cell automatically fills with sheath fluid at a controlled rate to prevent bubble
formation or entrapment. The STANDBY button turns amber after completion.
4. When the STANDBYlight is on, install a test tube of DIwater or leave the HTSconnected to the SIT.
5. Press the RUN fluid control button and run for 10 seconds.
6. Press the PRIMEbutton immediately followed by the RUNbutton.
7. Repeat step 6 ten times.
If more than 2 seconds pass between PRIMEand RUN in step 6, restart the procedure.
8. Install a 12 × 75 mm tube with 1mLof DIwater on the SIPand place the support arm under the tube.
Leave the cytometer in standby mode.
More information
l
Cytometer troubleshooting (page 104)
Adding a bead lot number
You need to be an administrator to perform this procedure.
Do not import a bead lot file as described in the BD
®
Cytometer Setup and Tracking Application Guide as the
standard bead lot files are not applicable to the BDFACSymphony™A5SE flow cytometer. Use the following
procedure to add a bead lot number instead.
Procedure
To add a bead lot number:
1. Select Cytometer >CST.
2. In the CS&T workspace, select Tools > Bead Lots.
The Bead Lots dialog opens.
3. Click New.
4. Enter the 6-digit part number for the BD
®
Cytometer Setup and Tracking Beads.
5. Enter the bead lot number.
32 BD FACSymphony™ A5 SE User's Guide
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