ArticleJournal of visualized experiments : JoVE2025
Simultaneous Label-Free Autofluorescence Multi-Harmonic Microscopy.
Article in Journal of visualized experiments : JoVE, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 5 papers.
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Who cites it
5 citing papers in PubMed.
- Autofluorescence profiles define depot-specific adipocyte phenotypes in perivascular and non-perivascular adipose tissue.Adipocyte · 2026Article
- Detecting drug-induced nephrotoxicity using simultaneous label-free autofluorescence multiharmonic microscopy.Toxicological sciences : an official journal of the Society of Toxicology · 2026Article
- Label-free multimodal nonlinear microscopy enabled by an optical parametric generator.APL photonics · 2026Article
- On the importance of simultaneous label-free multimodal nonlinear optical imaging for biomedical applications.APL photonics · 2025Article
- Unified Vibrational and Multiphoton Label-Free Nonlinear Microscopy for Simultaneous Chemical and Structural Imaging.IEEE journal of selected topics in quantum electronics : a publication of the IEEE Lasers and Electro-optics SocietyArticle
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Authors and funding
5 authors.
Funding
Abstract
Nonlinear optical microscopy images biological samples by detecting signals from the nonlinear interaction of ultrashort laser pulses with endogenous molecules. This method allows fast chemical and structural identification at subcellular resolution in a label or tag-free and nondestructive manner, thereby enabling a powerful approach to investigate cells and tissues. These distinctive nonlinear contrasts include multiphoton-excited autofluorescence and harmonic generation. Because each of these contrasts offers unique advantages and limitations, their combination and their spatiotemporal co-registration provide a complementary contrast palette that enhances the analytical capabilities of nonlinear optical microscopy. Therefore, our group developed Simultaneous Label-free Autofluorescence Multi-harmonic (SLAM) microscopy, an imaging technique that measures four or more concurrently generated nonlinear optical signals, aiming to identify distinct morphological, metabolic, and functional features in biological specimens. Here, we present a protocol for SLAM imaging of tissues, focusing on essential components of the technique, including the laser source, pulse compression, and microscope. In addition, we discuss sample preparation and outline the data processing pipeline for SLAM data. The presented workflow is suitable for investigating the metabolic state, arrangement, cellular responses, and composition of both human and animal tissues without relying on exogenous labels.
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Registered trials
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