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Simultaneous Label-Free Autofluorescence Multi-Harmonic Microscopy

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A deep tissue fluorescence imaging system with enhanced SHG detection capabilities.

Viera Crosignani¹, Sohail Jahid, Alexander S Dvornikov

¹Laboratory for Fluorescence Dynamics, Department of Biomedical Engineering, University of California Irvine, Irvine, California, 92697.

Microscopy Research and Technique

|March 11, 2014

Summary

A new two-photon fluorescence microscopy system offers high-quality second harmonic generation (SHG) imaging in challenging thick, turbid samples. Its innovative, large-area detection system excels at capturing forward-directed SHG signals.

Keywords:

deep tissue imaging second harmonic generation two-photon microscopy

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Area of Science:

Biomedical Optics
Microscopy Technology
Photonics

Background:

Second Harmonic Generation (SHG) microscopy is valuable for visualizing biological tissues.
Imaging thick, turbid samples with conventional microscopy is challenging due to light scattering.
SHG signals are typically forward-directed, complicating detection in transmission mode.

Purpose of the Study:

To introduce a novel two-photon fluorescence microscopy system for high-quality SHG imaging.
To demonstrate the system's effectiveness in thick and turbid media.
To compare forward and epi-detection schemes for SHG signal acquisition.

Main Methods:

Development of a novel two-photon fluorescence microscopy system.
Implementation of an innovative detection system with a large surface area.
Utilizing a transmission detection geometry optimized for forward-directed SHG signals.
Simultaneous data acquisition using both forward and epi-detection schemes on the same samples.

Main Results:

The novel system produces high-quality SHG images in thick, turbid media.
The large-area detection system effectively captures SHG photons.
Comparative data demonstrates the advantages of the transmission detection geometry for SHG.
The system is effective for both biological and artificial thick turbid samples.

Conclusions:

The developed two-photon microscopy system enables high-quality SHG imaging in challenging samples.
The innovative detection system and transmission geometry are key to its performance.
This technology offers a significant advancement for SHG microscopy of thick tissues.