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Speaker: Dr. Manjunath G. Javoor, PhD
Florian Schur & Michael Sixt groups
Institute of Science and Technology
Austria (ISTA), Austria
Title: "A molecular atlas of cellular actin networks at
single-filament resolution using montage cryo-
electron tomography."
Day and Date: Friday, September 18, 2026
Time: 12.00 Noon.
Venue: Room no. 350, Chemistry Department
Second floor, Annex
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Hosted by Prof. Ruchi Anand
Abstract Cell motility depends on the protrusive force generated by the actin
cytoskeleton at the leading edge of cells. In lamellipodia, thin
protrusions at the front of migrating cells, actin forms higher-order
networks with versatile physical properties. Previous studies have
hinted at the presence of different sub-populations of filaments and
their spatial organization within actin filament networks at the leading
edge of migrating cells. Due to limitations of existing experimental
methods, it has not been possible to fully describe the complexity of
the actin filament populations involved in protrusions at the leading
edge.
Cryo-electron tomography (cryo-ET) has been used to describe the actin
networks in cellular protrusions at single filament resolution in 3D.
However, these descriptions have been limited to 1-2 μm2 area of the
protrusions, providing merely snapshots of much larger assemblies that
span serval tens to hundreds of square microns area. Consequently, they
have not provided a holistic overview of the existing variability within
actin networks. We therefore lack a comprehensive understanding of how
actin network geometries orchestrate cell migration.
To address these challenges, I developed a montage cryo-electron
tomography workflow to obtain three-dimensional views of lamellipodial
regions covering areas of up to 100 μm2 at single-filament resolution.
Seamless three-dimensional tomogram stitching is achieved by combining
neural-network-based denoising strategies with novel optical-flow-based
algorithms, thereby preserving filament continuity across large
reconstructed volumes. Our computational analysis pipeline enabled us to
vectorize all filaments within these volumes, including filaments
reaching lengths of up to 4 μm. The data generated here represent a
valuable resource for the actin community, as they provide one of the
most detailed and extensive three-dimensional descriptions of actin
filament architecture in the lamellipodium. Specifically, they reveal
F-actin concentrations, filament length distributions, connectivity, and
the spatial organization of filament sub-populations. These datasets can
be used to support and constrain biophysical and theoretical models of
actin network organization, dynamics, and force generation.