Pathology Flow Cytometry Core Laboratory

If the LSRII isn't running right, try these corrective actions, but ONLY these.  Please do not do anything else to the machine without calling me first.


Possible Cause

Try this

Light scatter populations shifted dramatically, either at beginning of run or suddenly in middle of run

Air bubbles

Check level of sheath in sheath tank. Fill as necessary.  Push RUN.  Wait 10 min. Push PRIME.  When STANDBY button lights, push RUN.  Wait 60 seconds.  Prime HTS once.  Try running cells again.  If still bad, call Peter.  

Sheath tank empty

Call Peter.  Running the sheath tank dry requires about an hour’s worth of priming and refilling the system with sheath.

No events (sudden)


Check the paper towel where the “telltale” tube comes out.  See picture.  If wet, likely a clog.  Call Peter.  Don’t try to remedy on your own.  Priming HTS when it’s clogged could lead to damaged parts.

Few or no events when you first start running samples

Didn’t prime first?

Prime HTS 2x and prime flow cell 2x.  Re-run sample.

FACSDiva software is confined to one monitor


Position your mouse pointer at the rightmost edge of the FACSDiva window and slowly drag it to the right.  Should spread the Diva display across both monitors.

FACSDiva software doesn’t respond after acquiring first samples

System slow to save data

Sometimes after you acquire your first well the computer encounters some kind of trouble saving it.  You will know this is the problem because your well border will be green but there will be no floppy disc icon in it yet.  Wait 1-3 minutes.  The floppy disc icon should eventually appear and everything should be back to normal.

FACSDiva software freezes, not due to first sample pause

Software crash

Relaunch FACSDiva.  May have to shut down computer & restart.

Intermittent “odd” sample data patterns

Tube/plate selector switch set to tube

Move selector switch to plate.  See picture.  (Rare problem but it has happened.)