Multiphoton fluorescence microscopy: behavior of biological specimens under high-intensity illumination

Ping Chin Cheng; Bai-Ling Lin; Fu-Jen Kao; Chi-Kuang Sun

doi:10.1117/12.390536

4 July 2000 Multiphoton fluorescence microscopy: behavior of biological specimens under high-intensity illumination

Ping Chin Cheng, Bai-Ling Lin, Fu-Jen Kao, Chi-Kuang Sun

Proceedings Volume 4082, Optical Sensing, Imaging, and Manipulation for Biological and Biomedical Applications; (2000) https://doi.org/10.1117/12.390536
Event: Photonics Taiwan, 2000, Taipei, Taiwan

Abstract

Recent development in multi-photon fluorescence microscopy, second and third harmonic generation microscopy (SHG and THG) and CARS open new dimensions in biological studies. Not only the technologies allow probing the biological specimen both functionally and structurally with increasing spatial and temporal resolution, but also raise the interest in how biological specimens respond to high intensity illumination commonly used in these types of microscopy. We have used maize leaf protoplast as a model system to evaluate the photo-induced response of living sample under high intensity illumination. It was found that cells can be seriously damaged by high intensity NIR irradiation even the linear absorption coefficient in low in these wavelengths. Micro-spectroscopy of single chloroplast also allows us to gain insight on the possible photo-damage mechanism. In addition to fluorescence emission, second harmonic generation was observed in the maize protoplasts.

Citation Download Citation

Ping Chin Cheng, Bai-Ling Lin, Fu-Jen Kao, and Chi-Kuang Sun "Multiphoton fluorescence microscopy: behavior of biological specimens under high-intensity illumination", Proc. SPIE 4082, Optical Sensing, Imaging, and Manipulation for Biological and Biomedical Applications, (4 July 2000); https://doi.org/10.1117/12.390536

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