Immunofluorescent flow cytometry inN dimensions

Abstract

The problem of the simultaneous use in flow cytometry of N greater than 2 antibodies in conjunction with two fluorochromes was investigated. Theoretical analysis led to a labeling procedure and reconstruction formula that allow N-dimensional labeling distributions to be obtained from two-dimensional fluorescence distributions. The general problem of M greater than or equal to 2 fluorochromes and N greater than M antibodies was shown to be reducible to the case of two fluorochromes. The method was tested by a triple labeling analysis of murine thymocytes.

Properties of human blood monocytes. I. CD91 expression and log orthogonal light scatter provide a robust method to identify monocytes that is more accurate than CD14 expression

“…With regard to SSC, it is somewhat unconventional to collect and display log SSC signals so the rationale will be discussed in more detail below. Some antibody combinations were “multiplexed” with the same fluorochrome on more than one antibody, for example, . In these cases, the different populations labeled with the same fluorochrome could be distinguished by SSC differences, by differences in the fluorescence intensity of various populations and/or by differential labeling with another antibody.…”

Small-Volume Flow Cytometry-Based Multiplex Analysis of the Activity of Small GTPases

“…1 ) [ 17 , 20 ]. Flow cytometry is capable of exciting at multiple absorption bands and detecting fluorescence at different emission wavelengths, and it is then possible to detect various analytes, simultaneously from a single sample [ 21 – 25 ]. The commercially available cytoplex beads used by us have up to 12 different intensity levels that can be used as unique bead identifiers.…”