title: Targeted sorting of single virus-infected cells of the coccolithophore Emiliania huxleyi.
abstract: Discriminating infected from healthy cells is the first step to understanding the mechanisms and ecological implications of viral infection. We have developed a method for detecting, sorting, and performing molecular analysis of individual, infected cells of the important microalga Emiliania huxleyi, based on known physiological responses to viral infection. Of three fluorescent dyes tested, FM 1-43 (for detecting membrane blebbing) gave the most unequivocal and earliest separation of cells. Furthermore, we were able to amplify the genomes of single infected cells using Multiple Displacement Amplification. This novel method to reliably discriminate infected from healthy cells in cultures will allow researchers to answer numerous questions regarding the mechanisms and implications of viral infection of E. huxleyi. The method may be transferable to other virus-host systems.
authors.affiliation: Bigelow Laboratory for Ocean Sciences, West Boothbay Harbor, Maine, United States of America. jmartinez@bigelow.org
authors.forename: Joaquín
authors.initials: J
authors.lastname: Martínez Martínez
authors.forename: Nicole J
authors.initials: NJ
authors.lastname: Poulton
authors.forename: Ramunas
authors.initials: R
authors.lastname: Stepanauskas
authors.forename: Michael E
authors.initials: ME
authors.lastname: Sieracki
authors.forename: William H
authors.initials: WH
authors.lastname: Wilson
mesh_terms: D005434:Flow Cytometry / Q000379:methods*; D005456:Fluorescent Dyes / Q000378:metabolism; D058087:Haptophyta / Q000166:cytology* / Q000821:virology*; D017862:Phycodnaviridae / Q000502:physiology*; D016133:Polymerase Chain Reaction; D011506:Proteins / Q000378:metabolism
chemical_list: D005456:Fluorescent Dyes; D011506:Proteins