Cell Analysis

Our staff is experienced with the majority of flow cytometry protocols and is available to assist you with assay design, fluorochrome, and control choices for your assays.

Step by Step Guide

  1. Choose markers and fluorochromes.
    If you can, use fluorochromes that span the entire light spectrum. Use bright fluors such as PE and APC for low-density antigens and use the dimmer fluors such as FITC and APC-Cy7 for higher density antigens. 

    Here are some links to some spectra visualization tools:
  2. Choose an instrument and schedule an appointment in iLab
    If you're not sure which analyzer to choose, just ask us. Here's a tool to estimate how much time you will need.

  3. Titer your antibodies. 
    Antibodies should be titered each time you purchase a new lot. Frequently you will find that you need far less antibody than the recommended amount. Here are some quick tips for titering antibodies.

  4. Don't forget controls.
    These are especially important for compensation.

  5. Prepare and stain your cells.
    The advantage of flow cytometry is cell-by-cell analysis, so we need single-cell suspensions. Here are some tips for preparing cells.

  6. Create a sample worklist using our templates or Kaluza G offline.

    Choosing an Analyzer

    Beckman Coulter Gallios – 3 Laser (405nm, 488nm, 638nm), 10 Parameters

    • Sample Loading in 32-Tube Capacity Carousel of 5ml Tubes (12 x 75 mm)

     

    Beckman Coulter Gallios561 – 4 Laser (405nm, 488nm, 561nm, 638nm), 10 Parameters

    • Sample Loading in 32-Tube Capacity Carousel of 5ml Tubes (12 x 75 mm)

     

    Bio-Rad ZE5/YETI – 5 Laser (UV, 405nm, 488nm, 561nm, 640nm), 28 Parameters

    • Sample Loading in:
      • 40-Tube Capacity Rack of 5ml Tubes (12 x 75 mm)
      • Single Tube Loading of 5ml Tubes (12 x 75 mm) from STAT Tube Position
      • 24-Tube Capacity Rack of Microcentrifuge Tubes
      • 96-Well and 384-Well Micoplates

     

    Cytek Aurora (Spectral) – 5 Laser (UV, 405nm, 488nm, 561nm, 640nm), 64 Channel

    • Sample Loading in:
      • Single 5ml Tubes (12 x 75 mm)
      • 96-Well Plate Loader

     

    Fluidigm CyTOF Helios (Mass Cytometer)

    • Sample Loading in Single 5ml Tubes (12 x 75 mm)

    Additional Resources

    Flow Cytometry: A Basic Introduction

    Apoptosis

    Contact Us

    Mailing Address

    CU Cancer Center Flow Cytometry SR
    12801 E. 17th Avenue
    Mail Stop 8127
    Aurora, CO 80045

    Email

    cc.flowcyto@ucdenver.edu

    Phone

    p| 303-724-3145 (analysis rooms)

    p| 303-724-3144 (sorting room)