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Fluorescence Lifetime Macro Imager for Biomedical Applications

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Published on: April 7, 2023

TWINKLE: An open-source two-photon microscope for teaching and research.

Manuel Schottdorf^1,2, P Dylan Rich¹, E Mika Diamanti¹

¹Princeton Neuroscience Institute, Princeton University, Princeton, NJ, United States of America.

|February 13, 2025

Summary

We developed "Twinkle," an open-source two-photon microscopy system designed for neuroscience research and education. This easy-to-set-up microscope offers high performance, simplifying complex two-photon imaging for wider accessibility.

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Area of Science:

Neuroscience
Biophysics
Optical Engineering

Background:

Two-photon microscopy is crucial for neuroscience research but often involves complex, integrated commercial or homemade systems.
The complexity of existing systems hinders customization, troubleshooting, and user training on fundamental principles.

Purpose of the Study:

To introduce "Twinkle," an open-source, high-performance two-photon microscope.
To provide an accessible and easy-to-set-up instrument for both neuroscience education and research.
To demonstrate the utility of open scientific instrumentation.

Main Methods:

Designed and built a fully open two-photon microscope system named "Twinkle."
Integrated a large field of view (>1 mm) using a modern objective with a 3 mm working distance and 2-inch optics.
Utilized GaAsP photomultiplier tubes to optimize fluorescence signal detection.

Main Results:

The "Twinkle" microscope demonstrated high performance and ease of setup.
The system was successfully used as a teaching tool in multiple two-week workshops.
Scientific use cases were exemplified, showcasing the instrument's research capabilities.

Conclusions:

"Twinkle" offers a viable, open-source alternative to complex commercial two-photon microscopes.
The system effectively bridges the gap between education and research in neuroscience.
This work contributes to the growing field of open scientific instrumentation, promoting accessibility and customization.