Circumventing photodamage in live-cell microscopy.
Circumventing photodamage in live-cell microscopy.
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DOI：
10.1016/b978-0-12-407761-4.00023-3
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发表时间：
2013
影响因子：
--
通讯作者：
Khodjakov, Alexey
中科院分区：
文献类型：
--
作者：
Magidson, Valentin;Khodjakov, Alexey
荧光显微镜已成为细胞生物学中的重要工具。这项技术使研究人员能够可视化组织，细胞，单个细胞器和细胞内大分子组装的动态。不幸的是，荧光显微镜不是完全“非侵入性的”，因为激发荧光团所需的高强度激发光对活细胞具有固有的毒性。由过度照明引起的生理变化可导致伪影和异常反应。在本章中，我们将回顾导致光毒性的主要因素，并讨论规避光损伤的实际解决方案。这些解决方案包括正确选择图像采集参数、优化滤镜组、硬件同步以及使用智能照明来避免不必要的曝光。
Fluorescence microscopy has become an essential tool in cell biology. This technique allows researchers to visualize the dynamics of tissue, cells, individual organelles and macromolecular assemblies inside the cell. Unfortunately, fluorescence microscopy is not completely ‘non-invasive’ as the high-intensity excitation light required for excitation of fluorophores is inherently toxic for live cells. Physiological changes induced by excessive illumination can lead to artifacts and abnormal responses. In this chapter we review major factors that contribute to phototoxicity and discuss practical solutions for circumventing photodamage. These solutions include the proper choice of image acquisition parameters, optimization of filter sets, hardware synchronization, and the use of intelligent illumination to avoid unnecessary light exposure.