Self-Training

Basics Topics:

      For those with no experience in flow cytometry, of for those desiring an in-depth review, the on-line tutorial Introduction to Flow Cytometry is highly recommended.  The tutorial was prepared and is offered by Becton Dickinson to train new users in the theory and operation of flow cytometric analyzers and cell sorters.  The entire tutorial requires about 4 hours to complete but can easily be broken up into shorter sub-sessions at the user's discretion.  The same tutorial is also available for viewing on CD in the FACScan Analyzer Lab which is in room 603 of the Wintrobe Building.

 

Advanced Topics:

     Users who are familiar with the operation of the FACScan but desire some help on more specialized topics should consider some of the following sources.

    Compensation.  A process that involves the electronic correction of fluorescence emissions which overlap (or spill over) into another fluorochrome's detector.  An excellent on-line discussion of the issue and a tutorial, prepared by Mario Roederer at the NIH Aids Vaccine Trial Laboratory, is available at http://www.drmr.com/compensation/.

    Conjugation of monoclonal antibodies.  For those who wish to prepare their own antibody conjugates, another website prepared by Mario Roederer covers the basics at http://www.drmr.com/abcon/.

    Data analysis and post-acquisition compensation.  As if compensation weren't confusing enough, with the increased numbers of  fluorochromes being used it is becoming more difficult and time consuming to set up compensation at the time of data acquisition.  Each fluorochrome must be paired with each of the other fluorochrome's emission spectra for compensation; if one is dealing with five, six or more colors the number of pairings (and interactions) becomes unmanageable.  The solution has been to do compensation in software after the data are collected.  Two recent publications discuss this issue and describe a new way of displaying compensated data and are available in the following pdf documents:

  1. Interpreting flow cytometry data: a guide for the perplexed.  Nature Immunol 7:681-685 (2006)
  2. HyperLog - A flexible log-like transform for negative, zero and positive valued data.  Cytometry 64A:34-42 (2005)

    The John Curtin Scholl of Medical Research (JCSMR) in Australia, has several good tutorials on flow cytometry, data analysis, statistics in flow cytometry, FACScan operation and antibody titration.

    DNA/Cell Cycle Analysis.  Tutorials on flow cytometric analysis of DNA and Cell Cycle as well as aneuploid DNA detection are both available from the University of Florida Flow Cytometry webpage.