AlanFung:Protocol/FACScan: Difference between revisions
Jump to navigation
Jump to search
>Alan6017518 |
>Alan6017518 |
||
Line 37: | Line 37: | ||
*Place the test tube with MilliQ H2O into the sample inlet | *Place the test tube with MilliQ H2O into the sample inlet | ||
*Continue the filling function until tube is free of bubbles | *Continue the filling function until tube is free of bubbles | ||
CellQuest Setup | |||
Open template (saved on hard drive, for example: untitled/lo group/shawn/RCP-30-5-AA…) | |||
Or, create a new template and place plots. | |||
Set file name and path | |||
Under the Acquire menu, click “Parameter Description” | |||
create folder | |||
click on “file” button and set to “sample ID” | |||
click “OK” | |||
enter name into sample ID line |
Revision as of 16:08, 26 June 2009
FACScan (Becton Dickinson)
Cytometer Appearance (Bottom Panel)
------------- -------------- -------------- | Sheath | |.2um Sheath | | Waste | | Tank | |Filter | | Tank | ------------- -------------- --------------
Cytometer Setup
- Turn on both the cytometer and the computer
- Open up the bottom panel on the cytometer
- Make sure the system is pressurized
(Vent valve should be in the upwards position) (Sheath tank should be tight and non-movable) (No gap between the sheath tank and the top of the rack)
Purge bubbles trapped in the filter
- Prepare a small waster beaker located near the sink
- Squeeze the tube and pull white cap off from the tube coming out from the sheath filter
- Release the water with bubbles into the small waste beaker until no bubbles are seen
Removal of bubbles from flow cell
- Open up the middle panel on the cytometer, locate the flow cell (a small window with fluid in it)
- Swing the supporting arm to the side
- Take out tube containing MilliQ H2O
- Turn knob on the control panel to <Drain> in order to drain the flow cell (Notice the fluid level drops)
- Once the flow cell is drained, turn the knob to <Fill> (Notice the fluid level rises)
- Repeat the drain and fill procedure 3 times or until no bubbles are seen on the wall of the flow cell
- Leave the control knob on <Fill>
- Close the middle panel
Purge bubble from ? tube
- Prepare test tube with MilliQ H2O
- Open up the bottom panel
- Locate the tube linked to the orange valve
- Place the test tube with MilliQ H2O into the sample inlet
- Continue the filling function until tube is free of bubbles
CellQuest Setup Open template (saved on hard drive, for example: untitled/lo group/shawn/RCP-30-5-AA…) Or, create a new template and place plots. Set file name and path Under the Acquire menu, click “Parameter Description” create folder click on “file” button and set to “sample ID” click “OK” enter name into sample ID line