FidlTrack: high-fidelity structure-aware single particle tracking resolves intracellular molecular motion in...

Area of Science:

• Cellular and Molecular Biology
• Biophysics
• Biotechnology

Background:

• Single Particle Tracking (SPT) is crucial for studying macromolecule dynamics in live cells.
• Current SPT methods struggle with high particle velocities/densities and lack reliability assessment in subcellular spaces.

Purpose of the Study:

• To introduce FidlTrack, a novel methodology to benchmark and enhance SPT fidelity.
• To enable reliable tracking of macromolecule dynamics in complex subcellular environments.

Main Methods:

• FidlTrack employs a parameter optimizer using synthetic data for fidelity maximization.
• Structure-aware tracking utilizes organelle information to constrain particle tracking.
• A tracking quality evaluator identifies and removes ambiguous track segments.

Main Results:

• FidlTrack significantly improves SPT fidelity, yielding up to a 2-fold enrichment in accurate data.
• Successfully applied to track proteins in the cytosol, mitochondria, and endoplasmic reticulum (ER).
• Demonstrated efficacy in analyzing ER protein dynamics, BACE1 cleavage, and intrabody binding.

Conclusions:

• FidlTrack offers a universal, open-access platform for improving SPT pipelines.
• Enables rational design of SPT experiments and resolves motion in challenging organelles.
• Provides enhanced accuracy for studying molecular dynamics in cellular research.