Abstract
Single-molecule localization microscopy (SMLM) enables the visualization of cellular nanostructures in vitro with sub-20 nm resolution. While substructures can generally be imaged with SMLM, the structural understanding of the images remains elusive. To better understand the link between SMLM images and the underlying structure, we developed a Monte Carlo (MC) simulation based on experimental imaging parameters and geometric information to generate synthetic SMLM imag-es. We chose the nuclear pore complex (NPC), a nanosized channel on the nuclear membrane which gates nucleo-cytoplasmic transport of biomolecules, as a test geometry for testing our MC model. Using the MC model to simulate SMLM images, we first optimized our clustering algorithm to separate >10^6 molecular localizations of fluorescently labeled NPC proteins into hundreds of individual NPCs in each cell. We then illustrated using our MC model to generate cellular sub-structures with different angles of labeling to inform our structural understanding through SMLM images obtained.
Citation
Wei-Hong Yeo, Yang Zhang, Amy E. Neely, Xiaomin Bao, Cheng Sun, and Hao F. Zhang. 2023. “Investigating uncertainties in single-molecule localization microscopy using experimentally informed Monte-Carlo simulation.” Nano Letters 23 (16): 7253-7259. https://doi.org/10.1021/acs.nanolett.3c00852.
@article{Yeo2023,
author = {Wei-Hong Yeo and Yang Zhang and Amy E. Neely and Xiaomin Bao and Cheng Sun and Hao F. Zhang},
doi = {10.1021/acs.nanolett.3c00852},
journal = {Nano Letters},
number = {16},
pages = {7253-7259},
title = {Investigating uncertainties in single-molecule localization microscopy using experimentally informed Monte-Carlo simulation},
volume = {23},
year = {2023}}