Icha Jaroslav, Weber Michael, Waters Jennifer C, Norden Caren
Max Planck Institute of Molecular Cell Biology and Genetics, Dresden, Germany.
Department of Cell Biology, Harvard Medical School, Boston, MA, USA.
Bioessays. 2017 Aug;39(8). doi: 10.1002/bies.201700003.
Phototoxicity frequently occurs during live fluorescence microscopy, and its consequences are often underestimated. Damage to cellular macromolecules upon excitation light illumination can impair sample physiology, and even lead to sample death. In this review, we explain how phototoxicity influences live samples, and we highlight that, besides the obvious effects of phototoxicity, there are often subtler consequences of illumination that are imperceptible when only the morphology of samples is examined. Such less apparent manifestations of phototoxicity are equally problematic, and can change the conclusions drawn from an experiment. Thus, limiting phototoxicity is a prerequisite for obtaining reproducible quantitative data on biological processes. We present strategies to reduce phototoxicity, e.g. limiting the illumination to the focal plane and suggest controls for phototoxicity effects. Overall, we argue that phototoxicity needs increased attention from researchers when designing experiments, and when evaluating research findings.
光毒性在实时荧光显微镜检查过程中经常发生，其后果往往被低估。激发光照射会对细胞大分子造成损伤，从而损害样品的生理功能，甚至导致样品死亡。在这篇综述中，我们解释了光毒性如何影响活样品，并强调除了光毒性的明显影响外，光照往往还有一些更微妙的后果，而这些后果在仅检查样品形态时是无法察觉的。光毒性的这些不太明显的表现同样存在问题，并且可能会改变从实验得出的结论。因此，限制光毒性是获取关于生物过程的可重复定量数据的先决条件。我们提出了减少光毒性的策略，例如将光照限制在焦平面，并建议对光毒性效应进行控制。总体而言，我们认为在设计实验和评估研究结果时，研究人员需要更加关注光毒性。
