Changes to lipid droplet configuration in mCMV-infected fibroblasts: live cell imaging with simultaneous CARS and two-photon fluorescence microscopy

Christine S. Y. Wong; Iain Robinson; Michael A. Ochsenkuehn; Jochen Arlt; William J. Hossack; Jason Crain

doi:10.1364/BOE.2.002504

Changes to lipid droplet configuration in mCMV-infected fibroblasts: live cell imaging with simultaneous CARS and two-photon fluorescence microscopy

Christine S. Y. Wong^*, Iain Robinson, Michael A. Ochsenkuehn, Jochen Arlt, William J. Hossack, Jason Crain

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

Abstract / Description of output

We have performed multimodal imaging of live fibroblast cells infected by murine cytomegalovirus (mCMV). The infection process was monitored by imaging the two-photon fluorescence signal from a GFP-expressing strain of mCMV, whilst changes to lipid droplet configuration were observed by CARS imaging. This allowed us to identify three visually distinct stages of infection. Quantitative analysis of lipid droplet number and size distributions were obtained from live cells, which showed significant perturbations across the different stages of infection. The CARS and two-photon images were acquired simultaneously and the experimental design allowed incorporation of an environmental control chamber to maintain cell viability. Photodamage to the live cell population was also assessed.

Original language	English
Pages (from-to)	2504-2516
Number of pages	13
Journal	Biomedical Optics Express
Volume	2
Issue number	9
DOIs	https://doi.org/10.1364/BOE.2.002504
Publication status	Published - 1 Sept 2011

Keywords / Materials (for Non-textual outputs)

Raman scattering microscopy
cytomegalovirus
microscopy
medical and biological imaging
cell analysis
nonlinear optics
four-wave mixing
fluorescence microscopy

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10.1364/BOE.2.002504

Changes to lipid droplet configuration in mCMV-infected fibroblasts live cell imaging with simultaneous CARS and two-photon fluorescence microscopy
This paper was published in Biomedical Optics Express and is made available as an electronic reprint with the permission of OSA. The paper can be found at the following URL on the OSA website: http://dx.doi.org/10.1364/BOE.2.002504. Systematic or multiple reproduction or distribution to multiple locations via electronic or other means is prohibited and is subject to penalties under law.
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title = "Changes to lipid droplet configuration in mCMV-infected fibroblasts: live cell imaging with simultaneous CARS and two-photon fluorescence microscopy",

abstract = "We have performed multimodal imaging of live fibroblast cells infected by murine cytomegalovirus (mCMV). The infection process was monitored by imaging the two-photon fluorescence signal from a GFP-expressing strain of mCMV, whilst changes to lipid droplet configuration were observed by CARS imaging. This allowed us to identify three visually distinct stages of infection. Quantitative analysis of lipid droplet number and size distributions were obtained from live cells, which showed significant perturbations across the different stages of infection. The CARS and two-photon images were acquired simultaneously and the experimental design allowed incorporation of an environmental control chamber to maintain cell viability. Photodamage to the live cell population was also assessed.",

keywords = "Raman scattering microscopy, cytomegalovirus, microscopy, medical and biological imaging, cell analysis, nonlinear optics, four-wave mixing, fluorescence microscopy",

author = "Wong, {Christine S. Y.} and Iain Robinson and Ochsenkuehn, {Michael A.} and Jochen Arlt and Hossack, {William J.} and Jason Crain",

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doi = "10.1364/BOE.2.002504",

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AU - Wong, Christine S. Y.

AU - Robinson, Iain

AU - Ochsenkuehn, Michael A.

AU - Arlt, Jochen

AU - Hossack, William J.

AU - Crain, Jason

PY - 2011/9/1

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AB - We have performed multimodal imaging of live fibroblast cells infected by murine cytomegalovirus (mCMV). The infection process was monitored by imaging the two-photon fluorescence signal from a GFP-expressing strain of mCMV, whilst changes to lipid droplet configuration were observed by CARS imaging. This allowed us to identify three visually distinct stages of infection. Quantitative analysis of lipid droplet number and size distributions were obtained from live cells, which showed significant perturbations across the different stages of infection. The CARS and two-photon images were acquired simultaneously and the experimental design allowed incorporation of an environmental control chamber to maintain cell viability. Photodamage to the live cell population was also assessed.

KW - Raman scattering microscopy

KW - cytomegalovirus

KW - microscopy

KW - medical and biological imaging

KW - cell analysis

KW - nonlinear optics

KW - four-wave mixing

KW - fluorescence microscopy

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Changes to lipid droplet configuration in mCMV-infected fibroblasts: live cell imaging with simultaneous CARS and two-photon fluorescence microscopy

Abstract / Description of output

Keywords / Materials (for Non-textual outputs)

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Intracellular imaging of host-pathogen interactions using combined CARS and two-photon fluorescence microscopies

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