Orbital Tracking Microscopy

Our lab has developed a state-of-the-art Orbital Tracking Microscopy (OTM) system for high-precision single-particle tracking in three dimensions. This system enables us to track dynamic processes with nanometer spatial resolution and millisecond temporal precision over extended time scales. By combining advanced optics, real-time feedback loops, and dual-color detection, our setup pushes the boundaries of single-particle tracking in living and dynamic systems.

Key Features of Orbital Tracking Microscopy

The OTM system is designed for robust and flexible tracking, integrating cutting-edge components and custom-built solutions:

• Rotating excitation laser: The system employs an orbiting laser to continuously scan around the tracked particle, ensuring high-resolution position updates.
• 3D tracking with active feedback: Real-time adjustments maintain the particle in focus, preventing loss of z-resolution during extended measurements.
• Dual-color tracking: Independent and correlative motion tracking of two spectrally distinct particles in real-time.
• Multi-modal spectroscopic capabilities: Simultaneous single-particle tracking and Förster Resonance Energy Transfer (FRET) measurements to probe molecular interactions.
• High temporal resolution: Response times as fast as 5 ms, allowing us to follow rapidly diffusing particles and dynamic cellular events.

Applications of Orbital Tracking Microscopy

This versatile setup has been applied across a range of biological and biophysical studies, including:

• Investigating molecular interactions: Simultaneous dual-color tracking and FRET analysis.
• Observing intracellular cargo transport: Endosome motion and mitochondrial dynamics.
• Characterizing diffusion and conformational changes: Biomolecules in solution.

This setup serves for advanced biophysical research, such as probing molecular interactions, conformational changes, and energy transfer mechanisms with nanometer resolution.

Technical Advancements

Our orbital tracking system incorporates several innovative advancements:

• Nanometer-scale spatial resolution: It achieves ~3 nm lateral (xy) and ~20 nm axial (z) precision.
• Extended tracking range: it is capable of monitoring particles over distances exceeding 100 μm and time scales up to 10 minutes.
• Real-time data acquisition and processing:A LabVIEW-controlled Field Programmable Gate Array (FPGA) ensures seamless data synchronization and storage.