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Journal of Biomedical Optics • Open Access

Wavelength-dependent backscattering measurements for quantitative real-time monitoring of apoptosis in living cells
Author(s): Christine S. Mulvey; Carly A. Sherwood; Irving J. Bigio

Paper Abstract

Apoptosis-programmed cell death-is a cellular process exhibiting distinct biochemical and morphological changes. An understanding of the early morphological changes that a cell undergoes during apoptosis can provide the opportunity to monitor apoptosis in tissue, yielding diagnostic and prognostic information. There is avid interest regarding the involvement of apoptosis in cancer. The initial response of a tumor to successful cancer treatment is often massive apoptosis. Current apoptosis detection methods require cell culture disruption. Our aim is to develop a nondisruptive optical method to monitor apoptosis in living cells and tissues. This would allow for real-time evaluation of apoptotic progression of the same cell culture over time without alteration. Elastic scattering spectroscopy (ESS) is used to monitor changes in light-scattering properties of cells in vitro due to apoptotic morphology changes. We develop a simple instrument capable of wavelength-resolved ESS measurements from cell cultures in the backward direction. Using Mie theory, we also develop an algorithm that extracts the size distribution of scatterers in the sample. The instrument and algorithm are validated with microsphere suspensions. For cell studies, Chinese hamster ovary (CHO) cells are cultured to confluence on plates and are rendered apoptotic with staurosporine. Backscattering measurements are performed on pairs of treated and control samples at a sequence of times up to 6-h post-treatment. Initial results indicate that ESS is capable of discriminating between treated and control samples as early as 10- to 15-min post-treatment, much earlier than is sensed by standard assays for apoptosis. Extracted size distributions from treated and control samples show a decrease in Rayleigh and 150-nm scatterers, relative to control samples, with a corresponding increase in 200-nm particles. Work continues to correlate these size distributions with underlying morphology. (Partial Abstract)

Paper Details

Date Published: 1 November 2009
PDF: 14 pages
J. Biomed. Opt. 14(6) 064013 doi: 10.1117/1.3259363
Published in: Journal of Biomedical Optics Volume 14, Issue 6
Show Author Affiliations
Christine S. Mulvey, Boston Univ. (United States)
Carly A. Sherwood, Boston Univ. (United States)
Irving J. Bigio, Boston Univ. (United States)

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