An open-source, high-resolution, automated fluorescence microscope

Area of Science:

• Biophysics
• Microscopy
• Open-source hardware

Background:

• Sophisticated fluorescence microscopes are essential for life sciences but expensive, creating a bottleneck in data production.
• Open-source hardware, leveraging 3D-printing and low-cost electronics, offers a potential solution.
• Existing low-cost options often compromise image quality, while advanced designs require significant expertise.

Purpose of the Study:

• To assemble an automated, high-resolution fluorescence microscope using open-source components and high-quality parts.
• To demonstrate the system's capability for high-resolution imaging of both fixed and live samples.
• To validate the system's performance for long-term live-cell imaging, single-molecule sensitivity, and super-resolution microscopy.

Main Methods:

• Integration of the UC2 microscopy toolbox with high-quality components, electronics, and software.
• Assembly of an automated, high-resolution fluorescence microscope.
• Testing with fixed and live biological samples, including long-term incubations and super-resolution experiments.

Main Results:

• Successful assembly of an automated, high-resolution fluorescence microscope.
• Demonstration of high-resolution imaging for fixed and live samples, including multi-day live-cell imaging.
• Achieved single-molecule sensitivity, enabling single particle tracking and SMLM super-resolution microscopy.

Conclusions:

• A low-budget, open-source fluorescence microscope can achieve high image quality and advanced capabilities comparable to commercial systems.
• The system empowers researchers with limited resources and serves as valuable educational material.
• High-quality scientific data generation is feasible for users with minimal technical expertise using affordable, open-source tools.