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A Protocol for Real-time 3D Single Particle Tracking

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Point-Spread Function Deformations Unlock 3D Localization Microscopy on Spherical Nanoparticles.

Teun A P M Huijben¹, Sarojini Mahajan², Masih Fahim¹

¹Department of Health Technology, Technical University of Denmark (DTU), Kongens Lyngby 2800, Denmark.

|October 16, 2024

Summary

This summary is machine-generated.

Researchers developed a new model to accurately image nanoparticle (NP) surface functionalization using super-resolution microscopy. This fast and precise method reveals NP surface coverage, crucial for applications like drug delivery and biosensing.

Keywords:

DNA-paint nanoparticles plasmonics point-spread function single-molecule localization microscopy surface functionalization

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Area of Science:

Nanotechnology
Biophysics
Microscopy

Background:

Nanoparticles (NPs) are vital in biosensing, drug delivery, and photothermal therapy.
Surface functionalization critically impacts NP performance.
Conventional super-resolution microscopy struggles with NP-induced artifacts in fluorophore localization.

Purpose of the Study:

To develop an accurate analytical point-spread function (PSF) model for fluorophores near spherical NPs.
To overcome systematic mislocalizations caused by NPs in super-resolution imaging.
To enable precise 3D localization of surface functional groups on NPs.

Main Methods:

Derived an analytical PSF model for a fluorophore near a spherical NP.
Achieved a four-orders-of-magnitude speedup compared to numerical methods.
Applied the model to DNA-PAINT super-resolution microscopy data of DNA-coated gold NPs.

Main Results:

Demonstrated <5 nm precision in extracting 3D positions of surface functional groups.
Revealed inhomogeneous surface coverage on NPs.
Validated the model's accuracy and speed for real-world applications.

Conclusions:

The developed analytical PSF model accurately corrects for NP-induced artifacts in super-resolution microscopy.
The method provides fast, precise, and accessible analysis of NP surface functionalization.
This approach is poised to become a standard for imaging NPs in nanomedicine and biosensing.