ArticleBiophysical journal1999
Ca2+ fluorescence imaging with pico- and femtosecond two-photon excitation: signal and photodamage.
Article in Biophysical journal, 1999. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 86 papers.
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Who cites it
86 citing papers in PubMed.
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- Simultaneous two- and three-photon multiplane imaging across cortical layers in freely moving mice.Nature methods · 2026Article
- Live-cell STED microscopy enables 50 nm resolution imaging with preserved cell proliferation.Scientific reports · 2026Article
- Gentle Label-Free Nonlinear Optical Imaging Relaxes Linear-Absorption-Mediated Triplet.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2025Article
- Photobleaching Analysis of Fluorescent Proteins in Two-Photon Microscopy at High Repetition Rates.Optics communications · 2025Article
- Optical sectioning methods in three-dimensional bioimaging.Light, science & applications · 2025Review
- Scanless two-photon voltage imaging.Nature communications · 2024Article
- More than double the fun with two-photon excitation microscopy.Communications biology · 2024Review
- Comparing confocal and two-photon CaBiophysical reports · 2023Review
- A three-photon head-mounted microscope for imaging all layers of visual cortex in freely moving mice.Nature methods · 2023Article
- All-optical interrogation of neural circuits in behaving mice.Nature protocols · 2022Review
- Tailored Multi-Color Dispersive Wave Formation in Quasi-Phase-Matched Exposed Core Fibers.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2022Article
- Article
- Multiphoton intravital microscopy of rodents.Nature reviews. Methods primers · 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
- Excitation light-induced phototoxicity during fluorescence imaging.Journal of biosciences · 2021Review
- Label-Free Multimodal Multiphoton Intravital Imaging.Advances in experimental medicine and biology · 2021Article
26 more citing papers are in PubMed but not listed here.
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Authors and funding
4 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The signal and limitations of calcium florescence imaging using nonresonant multiphoton absorption of near-infrared femto- and picosecond laser pulses were examined. The fluorescence changes of various Ca(2+)-indicators induced by transient increases of the intradendritic calcium concentration were evaluated by evoking physiological activity in neocortical neurons in rat brain slices. Photodamage was noticeable as irreversible changes in the parameters describing the calcium fluorescence transients. At higher two-photon excitation rates, a great variety of irregular functional and structural alterations occurred. Thus, signal and observation time were limited by phototoxic effects. At lower excitation rates, photodamage accumulated linearly with exposure time. Femtosecond and picosecond laser pulses were directly compared with respect to this cumulative photodamage. The variation of the pulse length at a constant two-photon excitation rate indicated that a two-photon excitation mechanism is mainly responsible for the cumulative photodamage within the investigated window of 75 fs to 3.2 ps. As a direct consequence, at low excitation rates, the same image quality is achieved irrespective of whether two-photon Ca(2+)-imaging is carried out with femto- or picosecond laser pulses.
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