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Related Experiment Video

Updated: Jun 3, 2026

Long-Term Imaging of Identified Neural Populations using Microprisms in Freely Moving and Head-Fixed Animals
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Long-Term Imaging of Identified Neural Populations using Microprisms in Freely Moving and Head-Fixed Animals

Published on: January 19, 2024

Label-free live brain imaging and targeted patching with third-harmonic generation microscopy.

Stefan Witte1, Adrian Negrean, Johannes C Lodder

  • 1Biophysics Group, Institute for Lasers, Life, and Biophotonics Amsterdam, VU University, De Boelelaan 1081, 1081 HV, Amsterdam, The Netherlands. s.m.witte@vu.nl

Proceedings of the National Academy of Sciences of the United States of America
|March 30, 2011
PubMed
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Researchers developed a new noninvasive method for imaging brain tissue using optical third-harmonic generation (THG). This label-free technique allows high-contrast visualization of neurons and other structures in live brain samples without fluorescent dyes.

Area of Science:

  • Neuroscience
  • Biomedical Optics
  • Medical Imaging

Background:

  • Visualizing neurons in living brain tissue is crucial for neuroscience and neurosurgery.
  • Current methods like two-photon laser-scanning microscopy (2PLSM) require fluorescent dyes, limiting noninvasive applications.
  • A noninvasive probe for brain morphology with micrometer resolution is highly desirable for optical biopsies.

Purpose of the Study:

  • To demonstrate high-contrast, label-free imaging of live brain tissue at cellular resolution.
  • To establish optical third-harmonic generation (THG) as a viable alternative to fluorescence-based contrast mechanisms.
  • To explore the application of THG imaging in guiding surgical tools within the brain.

Main Methods:

  • Utilized optical third-harmonic generation (THG) microscopy for imaging.

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  • Exploited the specific geometry and lipid content of brain tissue for partial phase matching of THG.
  • Applied THG imaging to visualize neurons, white matter, and blood vessels in live brain tissue.
  • Used THG-based imaging to guide micropipettes towards specific neurons.

    • Main Results:

    • Achieved high-contrast, label-free imaging of live brain tissue at cellular resolution.
    • Demonstrated simultaneous visualization of neurons, white-matter structures, and blood vessels.
    • Successfully guided micropipettes to target neurons using THG imaging.
    • Established THG as an effective contrast mechanism without fluorescent probes.

    Conclusions:

    • Optical third-harmonic generation (THG) enables rapid, noninvasive, label-free imaging of live brain tissue.
    • THG imaging offers a powerful alternative to fluorescence-based microscopy for neuroscience and diagnostics.
    • This technique paves the way for label-free microscopic live brain imaging and laser-guided microsurgery.