Analyzing cell structure and dynamics with confocal light scattering and absorption spectroscopic microscopy

Le Qiu; Edward Vitkin; Hui Fang; Munir M. Zaman; Charlotte Andersson; Saira Salahuddin; Mark D. Modell; Steven D. Freedman; Eugene B. Hanlon; Irving Itzkan; Lev T. Perelman

doi:10.1117/12.711694

7 February 2007 Analyzing cell structure and dynamics with confocal light scattering and absorption spectroscopic microscopy

Le Qiu, Edward Vitkin, Hui Fang, Munir M. Zaman, Charlotte Andersson, Saira Salahuddin, Mark D. Modell, Steven D. Freedman, Eugene B. Hanlon, Irving Itzkan, Lev T. Perelman

Author Affiliations +

Proceedings Volume 6436, Complex Dynamics and Fluctuations in Biomedical Photonics IV; 643606 (2007) https://doi.org/10.1117/12.711694
Event: SPIE BiOS, 2007, San Jose, California, United States

Abstract

We recently developed a new microscopic optical technique capable of noninvasive analysis of cell structure and cell dynamics on the submicron scale [1]. It combines confocal microscopy, a well-established high-resolution microscopic technique, with light scattering spectroscopy (LSS) and is called confocal light absorption and scattering spectroscopic (CLASS) microscopy. CLASS microscopy requires no exogenous labels and is capable of imaging and continuously monitoring individual viable cells, enabling the observation of cell and organelle functioning at scales on the order of 100 nm. To test the ability of CLASS microscopy to monitor cellular dynamics in vivo we performed experiments with human bronchial epithelial cells treated with DHA and undergoing apoptosis. The treated and untreated cells show not only clear differences in organelle spatial distribution but time sequencing experiments on a single cell show disappearance of certain types of organelles and change of the nuclear shape and density with the progression of apoptosis. In summary, CLASS microscopy provides an insight into metabolic processes within the cell and opens doors for the noninvasive real-time assessment of cellular dynamics. Noninvasive monitoring of cellular dynamics with CLASS microscopy can be used for a real-time dosimetry in a wide variety of medical and environmental applications that have no immediate observable outcome, such as photodynamic therapy, drug screening, and monitoring of toxins.

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Le Qiu, Edward Vitkin, Hui Fang, Munir M. Zaman, Charlotte Andersson, Saira Salahuddin, Mark D. Modell, Steven D. Freedman, Eugene B. Hanlon, Irving Itzkan, and Lev T. Perelman "Analyzing cell structure and dynamics with confocal light scattering and absorption spectroscopic microscopy", Proc. SPIE 6436, Complex Dynamics and Fluctuations in Biomedical Photonics IV, 643606 (7 February 2007); https://doi.org/10.1117/12.711694

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KEYWORDS

Light scattering

Microscopy

Confocal microscopy

Scattering

Spectroscopy

Particles

Beam splitters

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