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DOI:	10.1371/journal.pone.0104250
论文题名:	Mitigating Phototoxicity during Multiphoton Microscopy of Live Drosophila Embryos in the 1.0–1.2 µm Wavelength Range
作者:	Delphine Débarre; Nicolas Olivier; Willy Supatto; Emmanuel Beaurepaire
刊名:	PLOS ONE
ISSN:	1932-6203
出版年:	2014
发表日期:	2014-8-11
卷:	9, 期:8
语种:	英语
英文关键词:	Embryos ; Drosophila melanogaster ; Absorption ; Perturbation (geology) ; Fluorescence imaging ; Lasers ; Light pulses ; Zebrafish
英文摘要:	Light-induced toxicity is a fundamental bottleneck in microscopic imaging of live embryos. In this article, after a review of photodamage mechanisms in cells and tissues, we assess photo-perturbation under illumination conditions relevant for point-scanning multiphoton imaging of live Drosophila embryos. We use third-harmonic generation (THG) imaging of developmental processes in embryos excited by pulsed near-infrared light in the 1.0–1.2 µm range. We study the influence of imaging rate, wavelength, and pulse duration on the short-term and long-term perturbation of development and define criteria for safe imaging. We show that under illumination conditions typical for multiphoton imaging, photodamage in this system arises through 2- and/or 3-photon absorption processes and in a cumulative manner. Based on this analysis, we derive general guidelines for improving the signal-to-damage ratio in two-photon (2PEF/SHG) or THG imaging by adjusting the pulse duration and/or the imaging rate. Finally, we report label-free time-lapse 3D THG imaging of gastrulating Drosophila embryos with sampling appropriate for the visualisation of morphogenetic movements in wild-type and mutant embryos, and long-term multiharmonic (THG-SHG) imaging of development until hatching.
URL:	http://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0104250&type=printable
Citation statistics:
资源类型:	期刊论文
标识符:	http://119.78.100.158/handle/2HF3EXSE/18621
Appears in Collections:	过去全球变化的重建影响、适应和脆弱性科学计划与规划气候变化与战略全球变化的国际研究计划气候减缓与适应气候变化事实与影响

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File Name/ File Size	Content Type	Version	Access	License
journal.pone.0104250.PDF（1390KB）	期刊论文	作者接受稿	开放获取	CC BY-NC-SA	View Download

作者单位:

Laboratory for Optics and Biosciences, Ecole Polytechnique, CNRS UMR 7645, and INSERM U696, Palaiseau, France;Univ. Grenoble Alpes, LIPhy, Grenoble, France;CNRS, LIPhy, Grenoble, France;Laboratory for Optics and Biosciences, Ecole Polytechnique, CNRS UMR 7645, and INSERM U696, Palaiseau, France;Laboratory for Optics and Biosciences, Ecole Polytechnique, CNRS UMR 7645, and INSERM U696, Palaiseau, France;Laboratory for Optics and Biosciences, Ecole Polytechnique, CNRS UMR 7645, and INSERM U696, Palaiseau, France

Recommended Citation:

Delphine Débarre,Nicolas Olivier,Willy Supatto,et al. Mitigating Phototoxicity during Multiphoton Microscopy of Live Drosophila Embryos in the 1.0–1.2 µm Wavelength Range[J]. PLOS ONE,2014-01-01,9(8)

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