1 research outputs found
Investigating Uncertainties in Single-Molecule Localization Microscopy Using Experimentally Informed Monte Carlo Simulation
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