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Graphical analysis of flow cytometer data for characterizing controlled fluorescent protein display on λ phage
Author(s) -
Sokolenko Stanislav,
Nicastro Jessica,
Slavcev Roderick,
Aucoin Marc G.
Publication year - 2012
Publication title -
cytometry part a
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.316
H-Index - 90
eISSN - 1552-4930
pISSN - 1552-4922
DOI - 10.1002/cyto.a.22211
Subject(s) - phage display , fluorescence , flow cytometry , green fluorescent protein , biophysics , computational biology , biological system , biology , chemistry , nanotechnology , microbiology and biotechnology , materials science , biochemistry , physics , optics , peptide , gene
Abstract As native virus particles typically cannot be resolved using a flow cytometer, the general practice is to use fluorescent dyes to label the particles. In this work, an attempt was made to use a common commercial flow cytometer to characterize a phage display strategy that allows for controlled levels of protein display, in this case, eGFP. To achieve this characterization, a number of data processing steps were needed to ensure that the observed phenomena were indeed capturing differences in the phages produced. Phage display of eGFP resulted in altered side scatter and fluorescence profile, and sub‐populations could be identified within what would otherwise be considered uniform populations. Surprisingly, this study has found that side scatter may be used in the future to characterize the display of nonfluorescent proteins. © 2012 International Society for Advancement of Cytometry