title: Graphical analysis of flow cytometer data for characterizing controlled fluorescent protein display on λ phage.
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.
authors.affiliation: Department of Chemical Engineering, Waterloo Institute for Nanotechnology, University of Waterloo, Waterloo, Ontario N2L 3G1, Canada.
authors.forename: Stanislav
authors.initials: S
authors.lastname: Sokolenko
authors.forename: Jessica
authors.initials: J
authors.lastname: Nicastro
authors.forename: Roderick
authors.initials: R
authors.lastname: Slavcev
authors.forename: Marc G
authors.initials: MG
authors.lastname: Aucoin
mesh_terms: D010582:Bacteriophage lambda / Q000737:chemistry* / Q000235:genetics / Q000254:growth & development; D036022:Capsid Proteins / Q000737:chemistry / Q000235:genetics; D061505:Cell Surface Display Techniques / Q000379:methods*; D003196:Computer Graphics; D004926:Escherichia coli / Q000737:chemistry / Q000821:virology; D005434:Flow Cytometry / Q000379:methods*; D005453:Fluorescence; D049452:Green Fluorescent Proteins / Q000737:chemistry* / Q000235:genetics; D008856:Microscopy, Fluorescence; D010957:Plasmids / Q000737:chemistry / Q000235:genetics; D013696:Temperature
chemical_list: D036022:Capsid Proteins; C041745:D protein, Enterobacteria phage lambda; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins