ArticleBiomedical optics express2021
Two-photon peak molecular brightness spectra reveal long-wavelength enhancements of multiplexed imaging depth and photostability.
Article in Biomedical optics express, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 8 papers.
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Who cites it
8 citing papers in PubMed.
- Analytical model enabling the correction of absorption and scattering in dual-color ratiometric multiphoton microscopy of brain tissue.Neurophotonics · 2026Article
- Sub-50-femtosecond gain-managed amplified pulses enhance nonlinear ablation efficiency.Biomedical optics express · 2026Article
- High-throughput hyperspectral fluorescence imaging using a high-speed silicon photomultiplier array.Biomedical optics express · 2025Article
- hyperPICASSO: adapting the mutual-information-based unmixing technique PICASSO to hyperspectral datasets.Optics letters · 2025Article
- Photobleaching Analysis of Fluorescent Proteins in Two-Photon Microscopy at High Repetition Rates.Optics communications · 2025Article
- HCMV strain- and cell type-specific alterations in membrane contact sites point to the convergent regulation of organelle remodeling.Journal of virology · 2024Article
- Video-rate two-photon microendoscopy using second harmonic resonance fiber scanning.Biomedical optics express · 2024Article
- Denoising multiplexed microscopy images in n-dimensional spectral space.Biomedical optics express · 2022Article
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Authors and funding
2 authors.
Funding
Abstract
The broad use of two-photon microscopy has been enabled in part by Ti:Sapphire femtosecond lasers, which offer a wavelength-tunable source of pulsed excitation. Action spectra have thus been primarily reported for the tunable range of Ti:Sapphire lasers (∼700-1000 nm). However, longer wavelengths offer deeper imaging in tissue via reduced scattering and spectral dips in water absorption, and new generations of pulsed lasers offer wider tunable ranges. We present the peak molecular brightness spectra for eight Alexa Fluor dyes between 700-1300 nm as a first-order surrogate for action spectra measured with an unmodified commercial microscope, which reveal overlapping long-wavelength excitation peaks with potential for multiplexed excitation. We demonstrate simultaneous single-wavelength excitation of six spectrally overlapping fluorophores using either short (∼790 nm) or long (∼1090 nm) wavelengths, and that the newly characterized excitation peaks measured past 1000 nm offer improved photostability and enhanced fidelity of linear spectral unmixing at depth compared to shorter wavelengths.
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