AlanFung:Protocol/FACScan: Difference between revisions

From ZhangLabWiki
Jump to navigation Jump to search
>Alan6017518
No edit summary
>Alan6017518
Line 39: Line 39:


==CellQuest Setup==
==CellQuest Setup==
Open template (saved on hard drive, for example:  untitled/lo group/shawn/RCP-30-5-AA…)
*Open template (saved on hard drive, for example:  untitled/lo group/shawn/RCP-30-5-AA…)
Or, create a new template and place plots.
*Or, create a new template and place plots.
Set file name and path
*Set file name and path
Under the Acquire menu, click “Parameter Description”
*Under the Acquire menu, click “Parameter Description”
create folder
*create folder
click on “file” button and set to “sample ID”
*click on “file” button and set to “sample ID”
click “OK”
*click “OK”
enter name into sample ID line
*enter name into sample ID line

Revision as of 16:09, 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