ArticlePloS one2011
Deconstructing the late phase of vimentin assembly by total internal reflection fluorescence microscopy (TIRFM).
Article in PloS one, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 44 papers.
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Who cites it
44 citing papers in PubMed, 108 citations in OpenAlex.
- Keratin filaments and soluble vimentin interact with isoform specificity to organize cell signaling.Genes & development · 2026Article
- Vimentin Intermediate Filaments: A Paradigm Shift From Static Structure to Dynamic Cytoplasmic Network.BioEssays : news and reviews in molecular, cellular and developmental biology · 2026Review
- Intermediate Filaments: Structure-Mechanics Crosstalk.Sub-cellular biochemistry · 2026Review
- Continuous self-repair protects vimentin intermediate filaments from fragmentation.Proceedings of the National Academy of Sciences of the United States of America · 2025Article
- Vimentin filament transport and organization revealed by single-particle tracking and 3D FIB-SEM.The Journal of cell biology · 2025Article
- Scale-dependent interactions enable emergent microrheological stress response of actin-vimentin composites.Soft matter · 2024Article
- Transport and Organization of Individual Vimentin Filaments Within Dense Networks Revealed by Single Particle Tracking and 3D FIB-SEM.bioRxiv : the preprint server for biology · 2024Article
- Keratin isoform shifts modulate motility signals during wound healing.bioRxiv : the preprint server for biology · 2024Article
- Keratin filament mechanics and energy dissipation are determined by metal-like plasticity.Matter · 2023Article
- Regulation of neurofilament length and transport by a dynamic cycle of phospho-dependent polymer severing and annealing.Molecular biology of the cell · 2023Article
- Vimentin: from a cytoskeletal protein to a critical modulator of immune response and a target for infection.Frontiers in immunology · 2023Review
- The N-Terminal Part of the 1A Domain of Desmin Is a Hot Spot Region for Putative PathogenicCells · 2022Article
- Article
- Dual-wavelength stopped-flow analysis of the lateral and longitudinal assembly kinetics of vimentin.Biophysical journal · 2022Article
- Genetic Insights into Primary Restrictive Cardiomyopathy.Journal of clinical medicine · 2022Review
- A general mathematical model for the in vitro assembly dynamics of intermediate filament proteins.Biophysical journal · 2022Article
- Multiscale mechanics and temporal evolution of vimentin intermediate filament networks.Proceedings of the National Academy of Sciences of the United States of America · 2021Article
- Molecular Insight into the Regulation of Vimentin by Cysteine Modifications and Zinc Binding.Antioxidants (Basel, Switzerland) · 2021Article
- Vimentin intermediate filaments stabilize dynamic microtubules by direct interactions.Nature communications · 2021Article
- Turn-on mode diarylethenes for bioconjugation and fluorescence microscopy of cellular structures.Proceedings of the National Academy of Sciences of the United States of America · 2021Article
Corrections and comments
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Authors and funding
8 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Quantitative imaging of intermediate filaments (IF) during the advanced phase of the assembly process is technically difficult, since the structures are several µm long and therefore they exceed the field of view of many electron (EM) or atomic force microscopy (AFM) techniques. Thereby quantitative studies become extremely laborious and time-consuming. To overcome these difficulties, we prepared fluorescently labeled vimentin for visualization by total internal reflection fluorescence microscopy (TIRFM). In order to investigate if the labeling influences the assembly properties of the protein, we first determined the association state of unlabeled vimentin mixed with increasing amounts of labeled vimentin under low ionic conditions by analytical ultracentrifugation. We found that bona fide tetrameric complexes were formed even when half of the vimentin was labeled. Moreover, we demonstrate by quantitative atomic force microscopy and electron microscopy that the morphology and the assembly properties of filaments were not affected when the fraction of labeled vimentin was below 10%. Using fast frame rates we observed the rapid deposition of fluorescently labeled IFs on glass supports by TIRFM in real time. By tracing their contours, we have calculated the persistence length of long immobilized vimentin IFs to 1 µm, a value that is identical to those determined for shorter unlabeled vimentin. These results indicate that the structural properties of the filaments were not affected significantly by the dye. Furthermore, in order to analyze the late elongation phase, we mixed long filaments containing either Alexa 488- or Alexa 647-labeled vimentin. The 'patchy' structure of the filaments obtained unambiguously showed the elongation of long IFs through direct end-to-end annealing of individual filaments.
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