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Updated: Mar 28, 2026

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

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

Published on: October 1, 2014

Note: Time-gated 3D single quantum dot tracking with simultaneous spinning disk imaging.

M S DeVore¹, D G Stich¹, A M Keller¹

¹Center for Integrated Nanotechnologies, Los Alamos National Laboratory, Mail Stop G755, Los Alamos, New Mexico 87545, USA.

The Review of Scientific Instruments

|January 3, 2016

Summary

This summary is machine-generated.

Recent microscope upgrades enable simultaneous 3D tracking and spinning disk imaging for cellular visualization. Photon time-gating enhances signal-to-noise, allowing time-resolved spectroscopy during single-particle tracking.

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Last Updated: Mar 28, 2026

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

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Published on: October 1, 2014

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Whole-cell Super-Resolution Imaging via DNA-PAINT on a Spinning Disk Confocal with Optical Photon Reassignment

Published on: January 6, 2026

Area of Science:

Biophysics
Microscopy
Cell Biology

Background:

Advanced microscopy techniques are crucial for visualizing cellular structures and molecular interactions.
Single-particle tracking (SPT) allows for the study of molecular dynamics within cells.
Limitations in current SPT methods include signal-to-noise ratios and limited temporal resolution.

Purpose of the Study:

To describe upgrades to a 3D tracking microscope.
To integrate simultaneous Nipkow spinning disk imaging with time-gated single-particle tracking (SPT).
To enable visualization of cellular environments around target molecules and perform time-resolved spectroscopy.

Main Methods:

Upgraded a 3D tracking microscope with Nipkow spinning disk imaging.
Incorporated photon time-gating into the SPT hardware.
Recorded single-photon arrival times for time-resolved measurements.

Main Results:

Achieved simultaneous 3D molecular tracking and spinning disk imaging.
Improved signal-to-noise ratio by discriminating against background noise and short-lived fluorescence.
Enabled time-resolved spectroscopy, including fluorescence lifetime and photon correlation measurements, during SPT.

Conclusions:

The upgraded microscope facilitates simultaneous visualization of cellular structures and molecular dynamics.
Photon time-gating significantly enhances the quality of SPT data.
The system allows for comprehensive, time-resolved characterization of single molecules in their cellular context.