Real-time molecular imaging of organelles in living cell by multifocus excitation CARS microscope

Takeo Minamikawa; Tsutomu Araki; Mamoru Hashimoto

doi:10.1117/12.842346

26 February 2010 Real-time molecular imaging of organelles in living cell by multifocus excitation CARS microscope

Takeo Minamikawa, Tsutomu Araki, Mamoru Hashimoto

Proceedings Volume 7569, Multiphoton Microscopy in the Biomedical Sciences X; 756927 (2010) https://doi.org/10.1117/12.842346
Event: SPIE BiOS, 2010, San Francisco, California, United States

Abstract

We demonstrated real-time imaging of organelles in a living HeLa cell using a multi-focus excitation CARS (coherent anti-Stokes Raman scattering) microscope. Chemical selective CARS imaging of lipids and proteins was demonstrated by observing CH2 and CH3 vibrations. Real-time imaging of lipid rich organelles such as the plasma membrane, mitochondria, and lipid rich vesicles was achieved by observing CH2 stretching vibrations of lipids. The image acquisition rate of 5 frames per second was achieved without any staining. We also demonstrated real-time CARS imaging of laser-induced disruption and reaction of organelles in a living HeLa cell. A near-infrared pulsed laser beam tightly focused on an organelle in a living cell produces ablation at the focal point, causing local disruption of the organelle. We visualized the spatial and temporal distributions of a lipid rich organelles in the cytoplasm of a living HeLa cell in laser-induced dissection. We also demonstrated real-time CARS imaging of disruption of a plasma membrane and its repair.

Citation Download Citation

Takeo Minamikawa, Tsutomu Araki, and Mamoru Hashimoto "Real-time molecular imaging of organelles in living cell by multifocus excitation CARS microscope", Proc. SPIE 7569, Multiphoton Microscopy in the Biomedical Sciences X, 756927 (26 February 2010); https://doi.org/10.1117/12.842346

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