lsrii troubleshooting tips
Download
Skip this Video
Download Presentation
LSRII Troubleshooting Tips

Loading in 2 Seconds...

play fullscreen
1 / 10

LSRII Troubleshooting Tips - PowerPoint PPT Presentation


  • 67 Views
  • Uploaded on

LSRII Troubleshooting Tips. Before starting each acquisition, check the following items: Waste tank is empty, but contains ~1 liter of bleach FACSFlow Sheath Fluid tank is full FACSFlow Sheath Fluid is connected correctly to instrument/reservoir No bubbles in the Sheath filter

loader
I am the owner, or an agent authorized to act on behalf of the owner, of the copyrighted work described.
capcha
Download Presentation

PowerPoint Slideshow about ' LSRII Troubleshooting Tips' - nicholas-madden


An Image/Link below is provided (as is) to download presentation

Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author.While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server.


- - - - - - - - - - - - - - - - - - - - - - - - - - E N D - - - - - - - - - - - - - - - - - - - - - - - - - -
Presentation Transcript
lsrii troubleshooting tips
LSRII Troubleshooting Tips
  • Before starting each acquisition, check the following items:
    • Waste tank is empty, but contains ~1 liter of bleach
    • FACSFlow Sheath Fluid tank is full
    • FACSFlow Sheath Fluid is connected correctly to instrument/reservoir
    • No bubbles in the Sheath filter
      • Tap the filter against a hard surface to move bubbles to the drain valve. A screwdriver is often useful to dislodge & move or “direct” bubbles to the drain line.
      • Drain the Sheath filter
      • Repeat process until all bubbles are removed from the Sheath filter
    • Check that there are no bubbles in Sheath line from Sheath filter into the side of the instrument
      • Drain the tubing line if necessary
lsrii troubleshooting tips1
LSRII Troubleshooting Tips
  • No events/Very few events
    • Restart the LSRII, FACSFlow Supply System, & Computer
      • Often there is a disconnect between the LSRII & the computer. A simple restart will sometimes fix this problem.
    • Has the correct tube been selected in FACSDiva?
      • Check that the correct tube has a Pointer next to it in FACSDiva.
    • Ensure that the Acquire Button has been selected on the Acquisition Control Panel.
    • Is tube pressurized?
      • Is LSRII in RUN mode?
        • Check that the GREEN RUN light is illuminated.
      • Is tube seated properly on SIP (Sample Injection Port)?
        • Wiggle tube to make sure tube is seated properly on support arm.
        • Ensure that tube is pushed up tightly against the Bal Seal.
        • Check that the Bal Seal is not worn. Change Bal seal if necessary.
      • Are you using the appropriate tube for the LSRII?
      • Is the tube cracked?
lsrii troubleshooting tips2
LSRII Troubleshooting Tips
  • No events/Very few events (continued)
    • Is the SIP (Sample Injection Port) clogged?
      • Perform Daily Clean (in RUN HIGH mode , ~15 minutes required)
        • 1 minute with 0.1M NaOH with support arm to side followed by

5 minutes of 0.1M with support arm under the tube

        • 1 minute with FACSRinse with support arm to side followed by

5 minutes of FACSRinse with support arm under the tube

        • 1 minute with dI H2O with support arm to side followed by 5 minutes

of dI H2O with support arm under the tube

        • Perform Daily Calibration. If improvement is seen, repeat until Daily Calibration

passes. If no improvement is seen, continue to next step.

      • Perform Aggressive Debubbling Procedure
        • Alternate between pressing the PRIME button and the RUN button

every 3 seconds.

          • This forces the Flow Cell to empty & refill very quickly with

FACSFlow Sheath Fluid.

        • Perform Daily Calibration. If improvement is seen, repeat until Daily Calibration

passes. If no improvement is seen, continue to next step.

lsrii troubleshooting tips3
LSRII Troubleshooting Tips
  • No events/Very few events (Continued)
      • Carefully insert a stylet into the Sample Probe
        • This will often dislodge a piece of debris or clump of cells from

the Sample Probe.

        • Perform Daily Calibration. If improvement is seen, repeat until Daily Calibration passes. If no improvement is seen, continue to next step.
      • Perform a Monthly Clean (in RUN HIGH mode, ~1.5 hours required)
        • Bypass/disconnect the Sheath filter
        • 45 minutes with 0.1M NaOH in Sheath tank and with 0.1M NaOH

in tube on SIP

        • 45 minutes with dI H2O in Sheath tank and with dI H2O in tube on SIP
        • 5 minutes with FACSFlow Sheath Fluid in Sheath tank & with dI H2O

in tube on SIP

        • Reconnect the Sheath filter & ensure that ALL bubbles are out of the Sheath filter before continuing
        • 5 minutes with FACSFlow Sheath Fluid in Sheath tank & with dI H2O

in tube on SIP

        • Perform Daily Calibration. If improvement is seen, repeat until Daily Calibration passes. If no improvement is seen, continue to next step.
lsrii troubleshooting tips4
LSRII Troubleshooting Tips
  • No events/Very few events (Continued)
      • Flush Flow Cell with syringe filled with 0.1M NaOH
        • Fill 10-20mL syringe with 0.1M NaOH & disconnect sheath line

from top of Flow Cell.

        • Ensure that an empty vessel is under SIP & tube is off SIP with support arm to side.
        • Connect syringe tubing to top of Flow Cell & force 0.1M NaOH into Flow Cell.
        • Repeat 3 times.
        • Reconnect sheath line.
        • Perform Daily Calibration. If improvement is seen, repeat until Daily Calibration passes. If no improvement is seen, continue to next step.
      • Check that the Blue Laser is not blocked with the cover.
      • Check that the HTS toggle switch is on TUBE & not on PLATE!
      • Call Service!!!
lsrii troubleshooting tips5
LSRII Troubleshooting Tips
  • High CV or Median is Outside Acceptable Range when Daily Calibration is performed
    • If Beads were not made fresh, prepare new Beads & re-run

Daily Calibration Assay

    • Check for air bubbles in the Sheath Filter & Sheath line
    • Check for air bubbles in Flow Cell
      • Perform Aggressive Debubbling (see Slide 4 for details)
    • Perform a Daily Clean (see Slide 4 for details)
    • Perform a Monthly Clean (See Slide 4 for details)
lsrii troubleshooting tips6
LSRII Troubleshooting Tips
  • Sample being aspirated very quickly from tube
    • Is the DCM (Droplet Containment Module) audible when a tube is on the SIP & the LSRII in RUN mode?
      • Yes - Move the support arm fully under the tube & ensure that tube is truly pressurized.
      • No - The DCM is Failing. Call BD service!!!
  • Tube is backfilling rapidly or SIP is dripping with support arm to side
    • Check that the SIP is not clogged (see Slide 4 for details)
    • Change the O-Ring
    • Call BD Service!!! - Possible clog in DCM tubing that must be repaired by a BDB Service Technician
  • For some samples that are known to clump (BAL, mono/mac, etc), filter fixed/stained samples using 12x75 filter top tubes prior to putting tube on SIP
lsrii troubleshooting tips7
LSRII Troubleshooting Tips
  • For some samples that are known to clump (BAL, mono/mac, etc), filter fixed/stained samples using 12x75 filter top tubes prior to putting tube on SIP.
lsrii spare parts to have on hand
LSRII Spare Parts to have on Hand
  • Sheath filter (BDB Catalog # 344678)
  • Bal seals (BDB Catalog # 343509)
  • Stylets (BDB Catalog # 343648)
  • O-Ring (BDB Catalog # 343615)
  • Tubing (ask your service engineer for extra tubing)
  • Screwdriver
lsrii reagents to have on hand
LSRII Reagents to have on Hand
  • FACSFlow Sheath Fluid (BDB Catalog # 342003)
  • FACSRinse (BDB Catalog # 340346)
  • D-PBS (Invitrogen/Gibco Catalog # 14190-144)
  • 8 Peak Rainbow Beads (Spherotech Catalog # RCP-30-5A)
  • 1X Rainbow Beads (Spherotech Catalog # RFP-30-5A)
  • FACSDiva 6.0 CST Beads (BDB Catalog # 641319)
  • FACSDiva 7.0+ CST Beads (BDB Catalog# 655051)
  • anti-Mouse Compensation Beads (BDB Catalog # 552843)
    • Use Negative Beads from kit for Daily Calibration
  • Sodium hydroxide (Sigma-Aldrich Catalog # S8045)
    • 1M NaOH pellets
  • dI H2O (any source is OK)
ad