Principles of Gating

James C. S. Wood1

1 Coulter Corporation, Miami, Florida
Publication Name:  Current Protocols in Cytometry
Unit Number:  Unit 1.8
DOI:  10.1002/0471142956.cy0108s03
Online Posting Date:  May, 2001
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Few aspects of flow cytometry are common to every application. Gating principles is one. This unit provides an excellent overview as well as detailed conceptual material on how gating works, when certain types of gating should or should not be applied, and the advantages and disadvantages of various techniques. This unit should be required reading for every technician using a flow cytometer and every graduate student starting out in the area, and is an excellent refresher for those more advanced in the technology. It brings out the historical development of our understanding of gating applications as well as providing a high level of technical accuracy.

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Table of Contents

  • Science of Gating
  • Mathematics to Combine Gates
  • Gating Strategies
  • Conclusion
  • Literature Cited
  • Figures
  • Tables
PDF or HTML at Wiley Online Library


PDF or HTML at Wiley Online Library



Literature Cited

   Auer, R.E., Starling, D., Weber, B., and Wood, J.C.S. 1993. A data acquisition system for flow cytometry with wide dynamic range analog to digital conversion and digital signal processing. Cytometry (Suppl.) 6:146A.
   Cupp, J.E., Leary, J.F., Cernichiari, E., Wood, J.C.S., and Doherty, R.A. 1984. Rare‐event analysis methods for detection of fetal red blood cells in maternal blood. Cytometry 5:138‐144.
   Frankel, D.S., Loken, M.R., Stelzer, G.T., Shults, K.E., and Bagwell, C.B. 1993. Neural network analysis of flow cytometric data for normal and leukemic bone marrow. Cytometry (Suppl.)6:44.
   Leary, J.F., Ellis, S.P., and McLaughlin, S.R. 1991. 3‐D autostereoscopic viewing of multidimensional data for principal component/biplot analysis and sorting. Cytometry (Suppl.)5:134‐135.
   Loken, M.R., Brosnan, J.M., Bach, B.A., and Ault, K.A. 1990. Establishing optimal lymphocyte gates for immunophenotyping by flow cytometry. Cytometry 11:453‐459.
   Mandy, F.F., Bergeron, M., Recktenwald, D., and Izaguirre, C.A. 1992. A simultaneous three‐color T cell subsets analysis with single laser flow cytometers using T cell gating protocol: Comparison with conventional two‐color immunophenotyping method. J Immunol. Methods 156:151‐156.
   McCoy, J.P. Jr., Chambers, W.H., Lakomy, R., Campbell, J.A., and Stewart, C.C. 1991. Sorting minor subpopulations of cells: Use of fluorescence as the triggering signal. Cytometry 12:268‐274.
   Murphy, R.F. 1985. Automated identification of subpopulations in flow cytometric list mode data using cluster analysis. Cytometry 6:302‐309.
   Nicholson, J.K.A., Jones, B.M., and Hubbard, M. 1993. CD4 T‐lymphocyte determinations on whole blood specimens using a single‐tube three‐color assay. Cytometry 14:685‐689.
   Redelman, D. 1993. Improved procedures for training neural networks to analyze flow cytometric data. Cytometry (Suppl.)6:43.
   Snow, C. and Bauer, K. 1994. Usefulness of DNA fluorescence ratioing in reducing DNA measurement artifacts caused by aggregates. Cytometry (Suppl.)7:50.
   van den Engh, G. and Stokdijk, W. 1989. Parallel processing data acquisition system for multilaser flow cytometry and cell sorting. Cytometry 10:289‐293.
Key Reference
   Shapiro, H.M. 1995. Practical Flow Cytometry, 3rd ed. Wiley‐Liss, New York.
  A resource for more examples of the use of gating in flow cytometry applications.
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