Dynamic analysis of pathogen-infected host cells using quantitative phase microscopy

• Published in: Journal of Biomedical Optics Vol. 16; no. 3; p. 036004
• Main Authors: Lee, Seungrag, Lee, Ji Yong, Park, Chang-Soo, Kim, Young Ran, Rhee, Joon Haeng, Kim, Dug Young
• Format: Journal Article
• Language: English
• Published: United States 01.03.2011
• Subjects: Animals; Cell Line; Cell Size; Genes, Bacterial; Host-Pathogen Interactions - physiology; Microscopy - instrumentation; Microscopy - methods; Microscopy, Interference; Microscopy, Phase-Contrast; Mutation; Optical Devices; Optical Phenomena; Rats; Vibrio vulnificus - genetics; Vibrio vulnificus - pathogenicity; Virulence - genetics
• ISSN: 1083-3668; 1560-2281
• DOI: 10.1117/1.3548882

We present the real-time quantitative analysis of -infected host cells using quantitative phase microscopy (QPM) based on interferometric techniques. This provides the ability to retrieve the phase or optical path-length distribution over the cell with nanometer path-length sensitivity from a single interferogram image. We have used QPM to study dynamic cell morphologic changes and to noninvasively quantify the cell volumes of rat basophilic leukemia RBL-2H3 cells infected with strains: wild type (MO6-24/O) and RtxA1 toxin mutant (CMM770). During the process of infection in RBL-2H3 cells, the dynamic changes of quantitative phase images, cell volumes, and areas were observed in real time using QPM. In contrast, dramatic changes were not detected in RBL-2H3 cells infected with the noncytotoxic RtxA1 toxin mutant. The results showed good correlation between QPM analysis and biochemical assays, such as lactate dehydrogenase assay or -hexosaminidase release assay. We suggest that QPM is a powerful quantitative method to study the dynamic process of host cells infected with pathogens in a noninvasive manner.