Texas Tech University

Guidelines for Using the Attune

Universal Guidelines for the Attune

 

Prior to beginning your experiment

  1. Check waste and fluid levels

    • If necessary, empty waste container and add bleach to its fill-to-line

    • Add fluids, as needed, to their appropriate containers. Do not exceed the maximum fill-to-line.

    • If necessary, run start-up procedure.

 

During the experiment

  1. Always monitor your data collection using the time parameter

    • A clog before the intercept results in a higher flow rate. A clog in the waste line slows the speed of the sample. Both, of which, results in loss of signal.

    • Should this occur, stop acquiring your data and run the Sanitize SIP function (see below). If the problem persists, notify the laboratory manager.

    • Not running your sample at top speed and using a more dilute sample will help. Also, filtering samples.

  2. Stay below 35,000 events/sec with 10% abort rate.

  3. Use speed of 12.5 µL when setting voltages and when running beads.

 

After the experiment

  1. Wipe the Instrument Sample Injection Port (SIP) using the “bleach wipes” then clean and sanitize the SIP and sample lines or the Auto Sampler SIP and sample lines.

  2. Empty waste container and, if necessary, run the shutdown procedure.

     

    Sanitize the SIP for the instrument:

    • On the Instrument ribbon, click Sanitize.

    • From the dropdown menu, select Sanitize Attune™ SIP.

      The Sanitize dialog box appears and provides instructions to perform the Sanitize Attune™ SIP procedure.

    • Click Next to initiate the Sanitize SIP procedure.

       

      Sanitize the SIP for the Auto Sampler:

    • On the Instrument ribbon, click Sanitize.

    • From the dropdown menu, select Auto Sampler SIP.

      The Sanitize dialog box appears and provides instructions to perform the Sanitize Auto Sampler SIP procedure.

    • Click Next to initiate the Sanitize SIP procedure.

Note: It is especially important to perform the Sanitize SIP function when running sticky samples, e.g., DNA stains or beads. Never use Acridine Orange in your samples. It is incredibly difficult to remove from the flow cell. A dirty flow cell can falsely shift the median of your negative population by 1-2 logs. KEEP IT CLEAN!!!

College of Arts and Sciences Microscopy