Abstract
Flow cytometry is a widely available technique for analyzing cell-surface protein expression. Data obtained from flow cytometry is frequently used to produce fluorescence intensity histograms. Comparison of histograms can be useful in the identification of unknown molecules and in the analysis of protein expression. In this study, we examined the combination of a new smoothing technique called SiZer with information theory to measure the difference between cytometry histograms. SiZer provides cross-bandwidth smoothing and allowed analysis in feature space. The new methods were tested on a panel of monoclonal antibodies raised against proteins expressed on peripheral blood lymphocytes and compared with previous methods. The findings suggest that comparing information content of histograms in feature space is effective and efficient for identifying antibodies with similar cell-surface binding patterns.
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- Bagwell C. B., Hudson J. L., Irvin G. L., 3rd Nonparametric flow cytometry analysis. J Histochem Cytochem. 1979 Jan;27(1):293–296. doi: 10.1177/27.1.374589. [DOI] [PubMed] [Google Scholar]
- Boddy L., Wilkins M. F., Morris C. W. Pattern recognition in flow cytometry. Cytometry. 2001 Jul 1;44(3):195–209. doi: 10.1002/1097-0320(20010701)44:3<195::aid-cyto1112>3.0.co;2-h. [DOI] [PubMed] [Google Scholar]
- Givan A. L. Principles of flow cytometry: an overview. Methods Cell Biol. 2001;63:19–50. doi: 10.1016/s0091-679x(01)63006-1. [DOI] [PubMed] [Google Scholar]
- Lampariello F., Aiello A. Complete mathematical modeling method for the analysis of immunofluorescence distributions composed of negative and weakly positive cells. Cytometry. 1998 Jul 1;32(3):241–254. doi: 10.1002/(sici)1097-0320(19980701)32:3<241::aid-cyto11>3.0.co;2-n. [DOI] [PubMed] [Google Scholar]
- Lampariello F. On the use of the Kolmogorov-Smirnov statistical test for immunofluorescence histogram comparison. Cytometry. 2000 Mar 1;39(3):179–188. doi: 10.1002/(SICI)1097-0320(20000301)39:3<179::AID-CYTO2>3.0.CO;2-I. [DOI] [PubMed] [Google Scholar]
- Parikh H. H., Li W. C., Ramanathan M. Evaluation of an alternative to the Kolmogorov-Smirnov test for flow cytometric histogram comparisons. J Immunol Methods. 1999 Oct 29;229(1-2):97–105. doi: 10.1016/s0022-1759(99)00108-8. [DOI] [PubMed] [Google Scholar]
- Watson J. V. A brief history of numbers and statistics with cytometric applications. Cytometry. 2001 Feb 15;46(1):1–22. doi: 10.1002/1097-0320(20010215)46:1<1::aid-cyto1032>3.0.co;2-3. [DOI] [PubMed] [Google Scholar]
- Watson J. V. Proof without prejudice revisited: immunofluorescence histogram analysis using cumulative frequency subtraction plus ratio analysis of means. Cytometry. 2001 Jan 1;43(1):55–68. doi: 10.1002/1097-0320(20010101)43:1<55::aid-cyto1019>3.0.co;2-t. [DOI] [PubMed] [Google Scholar]
- Wells W. M., 3rd, Viola P., Atsumi H., Nakajima S., Kikinis R. Multi-modal volume registration by maximization of mutual information. Med Image Anal. 1996 Mar;1(1):35–51. doi: 10.1016/s1361-8415(01)80004-9. [DOI] [PubMed] [Google Scholar]
- Young A. J., Marston W. L., Dessing M., Dudler L., Hein W. R. Distinct recirculating and non-recirculating B-lymphocyte pools in the peripheral blood are defined by coordinated expression of CD21 and L-selectin. Blood. 1997 Dec 15;90(12):4865–4875. [PubMed] [Google Scholar]
- Zeng Q., Young A. J., Boxwala A., Rawn J., Long W., Wand M., Salganik M., Milford E. L., Mentzer S. J., Greenes R. A. Molecular identification using flow cytometry histograms and information theory. Proc AMIA Symp. 2001:776–780. [PMC free article] [PubMed] [Google Scholar]