The K2: Open-source simultaneous triple-color TIRF microscope for live-cell and single-molecule imaging.

Area of Science:

• Biophysics
• Cell Biology
• Microscopy

Background:

• Single-molecule imaging provides crucial insights into biomolecular dynamics and cellular functions.
• High-speed, sensitive imaging requires specialized, often costly and complex, custom-built microscopes.
• Limited accessibility hinders broader adoption of advanced single-molecule imaging techniques.

Purpose of the Study:

• To present an open-source, simultaneous triple-color total internal reflection fluorescence (TIRF) microscope, named K2.
• To lower the barrier to entry for dynamic single-molecule imaging in live cells.
• To provide comprehensive documentation for building and customizing the K2 microscope.

Main Methods:

• Designed and constructed the K2 microscope, a modular, open-source TIRF system.
• Enabled simultaneous triple-color imaging for enhanced molecular tracking.
• Provided detailed building instructions, parts lists, and CAD models for reproducibility.

Main Results:

• The K2 microscope facilitates high-speed, sensitive detection of single biomolecules.
• Its open-source nature and modular design increase accessibility for researchers.
• The system is suitable for live-cell imaging and studying biomolecular interactions.

Conclusions:

• The K2 microscope democratizes advanced single-molecule imaging.
• Researchers can adapt the modular design for specific needs or integrate components into existing setups.
• This open-source approach accelerates research in cell biology and biophysics.