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A novel method for detection of virus‐infected cells through moving optical gradient fields using adenovirus as a model system
Author(s) -
Soo Hoo William,
Wang Mark,
Kohrumel Joshua R.,
Hall Jeff
Publication year - 2004
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.10097
Subject(s) - hela , green fluorescent protein , reporter gene , viral vector , cell culture , adenovirus infection , adenoviridae , biology , motility , virology , virus , microbiology and biotechnology , genetic enhancement , cell , gene expression , gene , recombinant dna , genetics , biochemistry
Background Most methods for cellular analysis require labeling with specific antibodies or dyes and are often destructive. We have developed a technology called Optophoresis™, which measures cell physiology based on the cell's motion in a near‐infrared optical gradient. This technique does not require labels, is nondestructive, and involves minimal sample processing. Methods We have used Optophoresis to interrogate nonproductive and productive adenovirus‐infected cell lines. Using an adenoviral vector containing green fluorescent protein (GFP) as a secondary assay, we show that viral infection can be monitored with Optophoresis. Results In HeLa cells, adenovirus infection after 24 h caused a 12% to 17% increase in optophoretic motility of the cells. In 293 cells, adenovirus infection resulted in a 40% increase in the optophoretic motility. The P values obtained were 4.5 × 10 −11 between noninfected and infected HeLa cells, and 2.1 × 10 −13 between noninfected and infected 293 cells. Cells infected with adenovirus lacking the GFP reporter gene gave similar shifts. In a time course, we observed an optophoretic shift after 4 h of infection, well before GFP expression. Conclusions Optophoresis provides nondestructive, label‐free analysis of viral infection. Detection is independent of reporter gene expression and can be observed early in the infection process. © 2004 Wiley‐Liss, Inc.