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Updated: Jun 20, 2025

Simultaneous Interference Reflection and Total Internal Reflection Fluorescence Microscopy for Imaging Dynamic Microtubules and Associated Proteins

Simultaneous Interference Reflection and Total Internal Reflection Fluorescence Microscopy for Imaging Dynamic Microtubules and Associated Proteins

Published on: May 3, 2022

Unified Simulation Platform for Interference Microscopy.

Felix Hitzelhammer^1,2,3, Anežka Dostálová^4,2,3, Ilia Zykov^2,3

¹Institute of Physics, University of Graz Universitätsplatz 5, 8010 Graz, Austria.

|July 22, 2024

Summary

This summary is machine-generated.

We developed a numerical toolbox to simulate interferometric scattering microscopy images. This tool accurately models various sample geometries and optical conditions, aiding in nanosensing applications.

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Area of Science:

Optical microscopy
Nanotechnology
Computational physics

Background:

Interferometric scattering microscopy (iSCAT) is vital for applications like mass photometry and particle tracking.
Accurate simulation of iSCAT is crucial for experimental design and data interpretation.

Purpose of the Study:

To present a versatile numerical toolbox for simulating images in interferometric scattering, coherent bright-field, and dark-field microscopy.
To validate the simulation accuracy against experimental data.

Main Methods:

Utilized the boundary element method (BEM) to calculate scattered fields for arbitrary sample geometries and substrate structures.
Employed a fully vectorial model to simulate the optical imaging setup.
Validated simulations with experimental results for diverse scatterer shapes and excitation angles.

Main Results:

Achieved excellent agreement between simulated and experimental microscopy images.
Observed significant contrast enhancement for scatterers near the Brewster angle.
Demonstrated the toolbox's capability to simulate complex sample geometries and layer structures.

Conclusions:

The developed numerical toolbox accurately simulates iSCAT, coherent bright-field, and dark-field microscopy images.
The observed Brewster angle contrast enhancement offers potential for advanced nanosensing applications.
The readily available MATLAB toolbox facilitates broader adoption and further research in optical microscopy simulations.