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University of Colorado Cancer Center

University of Colorado Cancer Center, A National Cancer Institute-designated Comprehensive Cancer Center
 

Cell Analysis

Guidelines


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


STEP BY STEP GUIDE

  1. Choose you 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. FluoroFinder is a great tool for finding reagents and choosing fluors.
  2. Choose an instrument and schedule your appointment. 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.
  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.