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3D Orbital Tracking in a Modified Two-photon Microscope: An Application to the Tracking of Intracellular Vesicles

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Parallel Three-Dimensional Tracking of Quantum Rods Using Polarization-Sensitive Spectroscopic Photon Localization

Biqin Dong^1,2, Brian T Soetikno¹, Xiangfan Chen²

¹Biomedical Engineering Department, Northwestern University, Evanston, Illinois 60208, United States.

|October 17, 2025

Summary

This summary is machine-generated.

We developed a new microscopy technique to track tiny semiconductor nanoparticles in 3D. This advanced imaging method precisely tracks quantum rods (QR) and their properties, enabling better biological research.

Keywords:

polarimetric imaging quantum rods single-particle tracking spectroscopy super-resolution microscopy

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Area of Science:

Biophysics
Nanotechnology
Optical Microscopy

Background:

Semiconductor nanocrystals, like quantum rods (QRs), are valuable fluorescent probes in biological research due to their brightness and photostability.
Their optical properties, including emission spectra and polarization, offer insights into mechanical properties and nanoenvironments.
Existing methods face limitations in simultaneously tracking multiple QRs in 3D with detailed spectral and polarization information.

Purpose of the Study:

To develop and validate a novel microscopy technique for high-precision, multi-parameter tracking of individual semiconductor nanocrystals.
To enable simultaneous acquisition of 3D position, fluorescence spectra, and polarization states of quantum rods.
To enhance the fidelity of parallel tracking of multiple quantum rods for studying complex biological dynamics.

Main Methods:

Development of a three-dimensional (3D), polarization-sensitive, spectroscopic photon localization microscopy (3D-Polar-SPLM).
Simultaneous 3D tracking of individual quantum rods (QRs) with high spatial, spectral, and polarization resolution.
Utilizing particle-specific spectral signatures for improved identification and tracking in heterogeneous populations.

Main Results:

Achieved lateral localization precision of 8 nm and axial precision of 35 nm for individual QRs.
Obtained spectral resolution of 2 nm and polarization angle measuring precision of 8 degrees.
Demonstrated improved tracking fidelity by using spectral profiles to distinguish individual QRs.

Conclusions:

The 3D-Polar-SPLM technology provides unprecedented capabilities for real-time, multi-parameter analysis of quantum rods.
This technique significantly enhances the ability to track multiple QRs in parallel, overcoming limitations of previous methods.
The developed microscopy offers new avenues for investigating real-time molecular dynamics in biological systems.