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Updated: Jan 17, 2026

Simultaneous Label-Free Autofluorescence Multi-Harmonic Microscopy
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Published on: August 29, 2025

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Simultaneous Label-Free Autofluorescence Multi-Harmonic Microscopy.

Kevin K D Tan1, Alejandro De la Cadena2, Edita Aksamitiene2

  • 1Beckman Institute for Advanced Science and Technology, University of Illinois Urbana-Champaign; Department of Bioengineering, University of Illinois Urbana-Champaign.

Journal of Visualized Experiments : Jove
|September 15, 2025
PubMed
Summary
This summary is machine-generated.

Simultaneous Label-free Autofluorescence Multi-harmonic (SLAM) microscopy images tissues using multiple nonlinear optical signals. This label-free technique reveals morphological, metabolic, and functional features in biological specimens without exogenous labels.

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Area of Science:

  • Biomedical Optics
  • Microscopy
  • Molecular Imaging

Background:

  • Nonlinear optical microscopy offers label-free, high-resolution imaging of biological samples.
  • Combining nonlinear contrasts enhances analytical capabilities for studying cells and tissues.
  • Existing methods have limitations in simultaneously acquiring multiple nonlinear signals.

Purpose of the Study:

  • To present a protocol for Simultaneous Label-free Autofluorescence Multi-harmonic (SLAM) microscopy of tissues.
  • To detail the essential components and workflow for SLAM imaging.
  • To enable comprehensive analysis of biological specimens without exogenous labels.

Main Methods:

  • SLAM microscopy utilizes ultrashort laser pulses to generate multiple nonlinear optical signals.
  • The technique involves specific laser sources, pulse compression, and microscope configurations.
  • Sample preparation and data processing pipelines are optimized for tissue imaging.

Main Results:

  • SLAM microscopy acquires four or more concurrent nonlinear optical signals.
  • The technique allows for the simultaneous measurement of autofluorescence and harmonic generation.
  • This provides a rich dataset for analyzing diverse biological features.

Conclusions:

  • SLAM microscopy enables label-free investigation of tissue morphology, metabolism, and function.
  • The presented protocol facilitates the application of SLAM imaging to human and animal tissues.
  • This advanced imaging approach enhances the understanding of biological specimens at subcellular resolution.