Abstract
Dynamic measurements of molecular machines can provide invaluable insights into their mechanism, but these measurements have been challenging in living cells. Here, we developed live-cell tracking of single fluorophores with nanometer spatial and millisecond temporal resolution in two and three dimensions using the recently introduced super-resolution technique MINFLUX. Using this approach, we resolved the precise stepping motion of the motor protein kinesin-1 as it walked on microtubules in living cells. Nanoscopic tracking of motors walking on the microtubules of fixed cells also enabled us to resolve the architecture of the microtubule cytoskeleton with protofilament resolution.
🔬 Techniques
✨ Fluorophores
🧪 Sample Preparation
🔬 Cell Lines
🏭 Microscope Brands
💾 Data Repositories
🏛️ Research Organizations (ROR)
Affiliated research institutions:
📊 Figures
Figure 1
MINFLUX tracking of kinesin-1 in fixed cells.
( A ) Kinesin-1 walks on microtubules (MTs) in a hand-over-hand manner. The apparent step size is 8 nm when the label is attached to the C-terminal tail domain and 16 nm when it is attached to the N-t...
Figure 2
MINFLUX tracking of kinesin-1 in live cells.
( A - D ) Tracking of full-length kinesin-1 labeled N-terminally with a HaloTag bound to JF646 in live U2OS cells. ( A ) Confocal images of GFP-u03b1-tubulin in untreated live U2OS cells, and overlaid...
Figure 3
3D MINFLUX tracking of kinesin-1.
( A , B ) 3D MINFLUX tracking. ( A ) A 3D donut beam probes the intensity at ( B ) seven three-dimensionally distributed positions around a fluorophore. ( C , D ) 3D tracking with motor-PAINT in fixed...
Figure images are served from the NIH/NLM PubMed Central Open Access Subset or Europe PMC; copyright remains with the publishers and authors.
💬 Discussion
0 commentsNo comments yet. Be the first to start a discussion!
Leave a Comment