ArticleJournal of fluorescence2015
Dependence of Two-Photon eGFP Bleaching on Femtosecond Pulse Spectral Amplitude and Phase.
Article in Journal of fluorescence, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 5 papers.
What it found
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
The trial behind it
Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
5 citing papers in PubMed, 8 citations in OpenAlex.
- Photobleaching Analysis of Fluorescent Proteins in Two-Photon Microscopy at High Repetition Rates.Optics communications · 2025Article
- Multiphoton Bleaching of Red Fluorescent Proteins and the Ways to Reduce It.International journal of molecular sciences · 2022Article
- High throughput instrument to screen fluorescent proteins under two-photon excitation.Biomedical optics express · 2020Article
- Excitation and emission dual-wavelength confocal metalens designed directly in the biological tissue environment for two-photon micro-endoscopy.Biomedical optics express · 2020Article
- Fiber-based tunable repetition rate source for deep tissue two-photon fluorescence microscopy.Biomedical optics express · 2018Article
Corrections and comments
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Authors and funding
5 authors at 2 institutions in 1 country.
Funding
Abstract
Photobleaching is a key limitation in two-photon imaging of fluorescent proteins with femtosecond pulsed excitation. We present measurements of the dependence of eGFP photobleaching on the spectral amplitude and phase of the pulses used. A strong dependence on the excitation wavelength was confirmed and measured over a 800-950 nm range. A fiber continuum light source and pulse shaping techniques were used to investigate photobleaching with broadband, 15 fs transform limited, pulses with differing spectral amplitude and phase. Narrow band pulses, >150 fs transform limited, typical of femtosecond laser sources used in two-photon imaging applications, were also investigated for their photobleaching dependence on pulse dispersion and bandwidth. The bleach rate for broadband pulses was found to be primarily determined by the second harmonic spectrum of the excitation light. On the other hand, for narrow band excitation pulses with similar center wavelengths improvement in bleach rate was found to be mostly dependent on reducing the pulse length. A simple model to predict the relative bleach rates for broadband pulses is presented and compared to the experimental data.
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Registered trials
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