ArticlePloS one2014
Mitigating phototoxicity during multiphoton microscopy of live Drosophila embryos in the 1.0-1.2 µm wavelength range.
Article in PloS one, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 35 papers.
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Who cites it
35 citing papers in PubMed.
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- Standardized measurements for monitoring and comparing multiphoton microscope systems.Nature protocols · 2025Review
- Gentle Label-Free Nonlinear Optical Imaging Relaxes Linear-Absorption-Mediated Triplet.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2025Article
- Multiphoton microscopy at a microwatt level via gain-managed nonlinear amplification and pulse-picking.Biomedical optics express · 2025Article
- Dynamic imaging of myelin pathology in physiologically preserved human brain tissue using third harmonic generation microscopy.PloS one · 2025Article
- Quantifying DNA damage following light sheet and confocal imaging of the mammalian embryo.Scientific reports · 2024Article
- Improved image contrast in nonlinear light-sheet fluorescence microscopy using iScientific reports · 2024Article
- More than double the fun with two-photon excitation microscopy.Communications biology · 2024Review
- EVATOM: an optical, label-free, machine learning assisted embryo health assessment tool.Communications biology · 2024Article
- Image restoration of degraded time-lapse microscopy data mediated by near-infrared imaging.Nature methods · 2024Article
- Standardised Measurements for Monitoring and Comparing Multiphoton Microscope Systems.bioRxiv : the preprint server for biology · 2024Article
- Axonal architecture of the mouse inner retina revealed by second harmonic generation.PNAS nexus · 2022Article
- Imaging Neutrophil Migration in the Mouse Skin to Investigate Subcellular Membrane Remodeling Under Physiological Conditions.Journal of visualized experiments : JoVE · 2022Article
- Article
- Low-invasive 5D visualization of mitotic progression by two-photon excitation spinning-disk confocal microscopy.Scientific reports · 2022Article
- Phototoxicity induced in living HeLa cells by focused femtosecond laser pulses: a data-driven approach.Biomedical optics express · 2021Article
- An adaptive microscope for the imaging of biological surfaces.Light, science & applications · 2021Article
- High-pulse-energy multiphoton imaging of neurons and oligodendrocytes in deep murine brain with a fiber laser.Scientific reports · 2021Article
- Infrared Excitation Induces Heating and Calcium Microdomain Hyperactivity in Cortical Astrocytes.Biophysical journal · 2020Article
- FastBiomedical optics express · 2020Article
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4 authors.
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Abstract
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.
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