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Updated: Mar 24, 2026

Simultaneous Label-Free Autofluorescence Multi-Harmonic Microscopy

Simultaneous Label-Free Autofluorescence Multi-Harmonic Microscopy

Published on: August 29, 2025

Fast interferometric second harmonic generation microscopy.

Stéphane Bancelin¹, Charles-André Couture¹, Katherine Légaré¹

¹Institut National de la Recherche Scientifique, Centre Énergie Matériaux Télécommunications (INRS-EMT); 1650 Boul. Lionel-Boulet, Varennes (QC), J3X 1S2, Canada.

Biomedical Optics Express

|March 16, 2016

Summary

This summary is machine-generated.

We developed a faster Interferometric Second Harmonic Generation (I-SHG) microscopy technique for studying biological tissue polarity. This new laser scanning method images collagen fibril polarity 40x faster than traditional methods.

Keywords:

(170.6935) Tissue characterization (180.4315) Nonlinear microscopy (190.2620) Harmonic generation and mixing (190.4180) Multiphoton processes (190.4710) Optical nonlinearities in organic materials

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A Multimodal Wide-Field Fourier-Transform Raman Microscope

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Area of Science:

Biomedical Optics
Microscopy Techniques
Materials Science

Background:

Non-centrosymmetric structures in biological tissues exhibit polarity crucial for function.
Interferometric Second Harmonic Generation (I-SHG) microscopy is a technique used to study these structures.
Standard I-SHG microscopy, while effective, can be time-consuming.

Purpose of the Study:

To implement and validate a fast Interferometric Second Harmonic Generation (I-SHG) microscopy method.
To assess the feasibility of using laser scanning systems for rapid I-SHG imaging.
To accurately determine the polarity of non-centrosymmetric structures in biological samples.

Main Methods:

Calibration of phase shifts in a laser scanning system using a quartz plate.
Compensation for spatially varying phase shifts.
Imaging of periodically poled lithium niobate as a model system.
Application of the fast I-SHG method to biological tissue (tendon).

Main Results:

Successful calibration and phase shift compensation were achieved.
The fast I-SHG microscopy accurately retrieved phase distribution in model samples.
Polarity of collagen fibrils in tendon was successfully recovered.
Imaging time was reduced by approximately 40 times compared to standard I-SHG.

Conclusions:

Fast I-SHG microscopy using a laser scanning system is a viable and significantly accelerated method for studying tissue polarity.
The technique provides comparable results to standard I-SHG but with vastly improved speed.
This advancement holds potential for more efficient analysis of biological structures.