Advanced real-time classification methods for flow cytometry data analysis and cell sorting

James F. Leary; Lisa M. Reece; James A. Hokanson; Judah I. Rosenblatt

doi:10.1117/12.468345

28 May 2002 Advanced real-time classification methods for flow cytometry data analysis and cell sorting

James F. Leary, Lisa M. Reece, James A. Hokanson, Judah I. Rosenblatt

Proceedings Volume 4622, Optical Diagnostics of Living Cells V; (2002) https://doi.org/10.1117/12.468345
Event: International Symposium on Biomedical Optics, 2002, San Jose, CA, United States

Abstract

While many flow cytometric data analysis and 'discovery' methods have been developed, few of these have been applied to the problem of separating out purified cell subpopulations by cell sorting. The fundamental problem is that the data analysis techniques have been performed using relatively slow computational methods that take far more time than is allowed by the sort decision on a cell sorter (typically less than a millisecond). Thus cell sorting, which is really a form of 'real-time data classification,' is usually done with few, if any, multivariate statistical tools used either in the sort decision or in the evaluation of the correctness of the classification. We have developed new multivariate data analysis and 'data discovery' methods that can be implemented for real-time data classification for cell sorting using linked lookup tables. One multivariate 'data discovery' method, 'subtractive clustering,' has been used to find which clusters of cells are different between two or more files (cell samples) and to help guide analysis or sort boundaries for these cell subpopulations. Multivariate statistical methods (e.g. principal component analysis or discriminant function analysis) were implemented in linked lookup tables to establish analysis/sort boundaries that include 'costs (or penalties) of misclassification. Costs of misclassification provided a measure of the quality of the analysis/sort boundary and were expressed in simple terms that describe the tradeoff between yield and purity.

Citation Download Citation

James F. Leary, Lisa M. Reece, James A. Hokanson, and Judah I. Rosenblatt "Advanced real-time classification methods for flow cytometry data analysis and cell sorting", Proc. SPIE 4622, Optical Diagnostics of Living Cells V, (28 May 2002); https://doi.org/10.1117/12.468345

ACCESS THE FULL ARTICLE

INSTITUTIONAL
Select your institution to access the SPIE Digital Library.

SELECT YOUR INSTITUTION

PERSONAL
Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.

PERSONAL SIGN IN

No SPIE Account? Create one

PURCHASE THIS CONTENT

SUBSCRIBE TO DIGITAL LIBRARY

50 downloads per 1-year subscription

Members: $195

Non-members: $335 ADD TO CART

25 downloads per 1 - year subscription

Members: $145

Non-members: $250 ADD TO CART

PURCHASE SINGLE ARTICLE

Includes PDF, HTML & Video, when available