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Simultaneous Label-Free Autofluorescence Multi-Harmonic Microscopy
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Fiber-based dual-focus time-demultiplexed second harmonic generation microscopy.

Sandro Heuke, Fisseha Bekele Legesse, Adrian Lorenz

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    We developed a dual-focus second harmonic generation (SHG) microscopy method using affordable fiber optics. This technique enables faster imaging of distinct areas by creating two synchronized beams, enhancing microscopy efficiency.

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    Area of Science:

    • Biomedical Optics
    • Microscopy Technologies
    • Nonlinear Optics

    Background:

    • Second Harmonic Generation (SHG) microscopy is a valuable nonlinear optical imaging technique.
    • Conventional SHG microscopy can be limited by imaging speed and field of view.
    • Advancements in laser technology and fiber optics offer opportunities for improved microscopy systems.

    Purpose of the Study:

    • To present a novel dual-focus second harmonic generation (SHG) microscopy approach.
    • To leverage stable, compact, and inexpensive fiber technology for enhanced SHG imaging.
    • To demonstrate a method for increasing imaging speed and flexibility within the field of view.

    Main Methods:

    • Utilized a fiber laser system with a portion of its output coupled into a 100m single-mode fiber.
    • Amplified the coupled light to generate two separately guided beams with time-alternating pulse trains.
    • Implemented sequential SHG detection to acquire two individual images within a single scan.

    Main Results:

    • Achieved a dual-focus SHG microscopy configuration based on fiber technology.
    • Demonstrated the capability to image distinct areas within the field of view simultaneously.
    • Obtained imaging at twice the repetition rate of the fiber laser, with potential for extension to multiple foci.

    Conclusions:

    • The developed dual-focus SHG microscopy approach offers a cost-effective and efficient solution.
    • This fiber-based technology enhances imaging speed and allows for imaging of separate regions.
    • The system is scalable to multiple foci, offering significant potential for advanced biological imaging applications.