Work

Optical sectioning in wide-field two-photon microscopy using temporal focusing and random illumination

Year: 2026

Type: article

Abstract: Temporal focusing (TF) is a key tool in wide-field two-photon excitation fluorescence microscopy for confining fluorescence excitation to a thin layer around the focal plane. TF is typically implemented with 100 femtosecond laser pulses and large microscope magnifications (>40). In this work, we demonstrate that performing TF with a random speckled illumination rather than a collimated beam significantly improves the optical sectioning for long pulses and small magnifications. We derive simple formulas of the optical sectioning as a function of speckle angular divergence, pulse bandwidth, and microscope parameters. Our approach paves the way for optically sectioned wide-field nonlinear imaging using >200 fs pulses and low magnifications enabling large fields of view.

Source: Optics Letters

DOI: 10.1364/ol.592916

Authors: Xiangyi Li, Federico Vernuccio, Michal Marynowski, Assia Benachir, Sandro Heuke, Hervé Rigneault, Anne Sentenac

Language: English

Topic: Advanced Fluorescence Microscopy Techniques

Subfield: Biophysics

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: green

Funders: Chan Zuckerberg Initiative, Institut National de la Santé et de la Recherche Médicale, HORIZON EUROPE Framework Programme

Awards: G659800116, G7465385040, G8589192729

OpenAlex ID: W7128793848

FWCI:

Cites: 15

Cited by:

Related to: