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Flow Cytometry Core Facility

AccuriC6

Accuri C6 is an "easy to use" flow cytometer which can be connected to an autosampler (48 and 96 well, and 24 tube rack, we do not have one). Accuri C6 is a digital PC based bench top analyser, 2 laser system. Standard fluorochrome in use are identical to FACS Calibur. Even more colors to choose from with selectable lasers and filter replacement. Available to trained users with online booking.

Specifications

  • 488 nm (blue) laser
  • 640 nm (red) laser

 

Four fluorescent emission detectors:

  • FL1 530+/-15 nm (FITC/GFP)
  • FL2 585+/-20 nm (PE/PI)
  • FL3 >670 nm (PerCP-Cy5.5, PE-Cy5, PE-Cy7)
  • FL4 675+/-12.5 nm (APC)

 

Interchangeable bandpass filters: 780/60, 610/20, 630/30

 

Use the standard 3 detectors on the blue laser and 1 on the red laser OR all 4 detectors on the blue laser OR 2 detectors on the blue and 2 on the red laser.

Event rate of up to 10,000 events/second.

 

Procedure

  • Sign log book
  • Fill the sheat fluid tank with MQ water
  • Empty the waste tank appropriately
  • Check the levels of the decontamination and cleaning fluid tanks. If less than half the fluid left, fill the tanks.
    • Cleaning solution. Decon 90. 1ml+199ml MQ water.
    • Decontamination solution. 1ml NaHCL (Sodium Hydrochloride)+199ml MQ water.
  • Ensure that there is a FACS tube with MQ water places on the sample introduction probe.
  • Turn on the Accuri C6 and the pc.
  • Open the program CFlow Plus.
  • Open a wash template under “Documents”.
  • Wash template is set to run MQ water for 2 minutes.
  • Run a new FACS tube with MQ water.
  • Ensure that the amount of event are few, if not was for another 2 minutes.
  • Open a new workspace or a template.
  • Set up the appropriate dotplot/histograms, set run rate to unlimited, or wated cell number in a specific gate.
  • Run samples on medium or high.
  • If no events are seen, or the sample collection rate is slower than expected. Stop running the sample. Remove the tube and put a empty tube on the probe. Push backflush.
  • Once finished, remove the FACS tube and wipe the probe with tissues. Run MQ water for 2 minutes.
  • Then continue to run your samples. Ensure the samples are not run dry.
  • After running the samples, place a FACS tube with 3ml of cleaning solution on the sip.
  • Either select a new well, or open the wash template
  • Run cleaning solution for 2 minutes
  • Then run a tube with clean MQ water for 2 minutes
  • If the plots are relatively empty of any events turn of the machine.
  • If thre is still a lot of events in the plot, wash for another 2 minutes.
  • If the machine is not going to be used later, turn the machine off.
  • If someone is going to use the machine within the next hr, leave the machine on.
  • Always leave a tube with MQ water on the sip.

Disposal/Spills/Incidents

(Also see HMS handbook for K2 regarding HMS routines)

  • Remove any spilled liquids with paper towels, dispose towels in the autoclave waste, then wipe the bench with 70% EtOH
  • Report any accidents
  • If there is skin contact with either the cleaning or decontamination solution, wash under running water.

Training

  • Inexperienced users can get training from Brith Bergum. Experienced users need to show competence in using the machine.

Relevant SDS

  • Cleaning solution. Decon 90.
  • Decontamination solution. NaHCL (Sodium Hydrochloride)

Reference