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 images. 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 >106 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 substructures with different angles of labeling to inform our structural understanding through the SMLM images obtained.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 7253-7259 |
| Number of pages | 7 |
| Journal | Nano letters |
| Volume | 23 |
| Issue number | 16 |
| DOIs | |
| State | Published - Aug 23 2023 |
Funding
We acknowledge the generous support from the National Institutes of Health (R21GM141675, R01EY026078, R01EY019949, R01GM140478, R01GM139151, R01GM143397, R01AR075015, and U54CA268084); National Science Foundation (CBET-1706642, CHE-1954430, and EFRI-1830969); and the American Cancer Society Research Scholar Grant (RSG-21-018-01-DDC). W. Y. thanks the Christine Enroth-Cugell Fellowship for Vision and Neuroscience at Northwestern University for their support.
Keywords
- Monte Carlo simulation
- Single-molecule localization microscopy
- image processing
- nuclear pore complex
ASJC Scopus subject areas
- General Chemistry
- Condensed Matter Physics
- Mechanical Engineering
- Bioengineering
- General Materials Science