ArticleeLife2018
An optimized method for 3D fluorescence co-localization applied to human kinetochore protein architecture.
Article in eLife, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
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Who cites it
14 citing papers in PubMed.
- A Molecular Recruitment Colocalization Platform for Visualizing Multi-Protein Interactions and Engineering Biomolecular Condensates in Living Cells.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2025Article
- One step 4× and 12× 3D-ExM enables robust super-resolution microscopy of nanoscale cellular structures.The Journal of cell biology · 2025Article
- Calibrating Fluorescence Microscopy With 3D-Speckler (3D Fluorescence Speckle Analyzer).Bio-protocol · 2024Article
- PyF2F: a robust and simplified fluorophore-to-fluorophore distance measurement tool for Protein interactions from Imaging Complexes after Translocation experiments.NAR genomics and bioinformatics · 2024Article
- Unraveling the kinetochore nanostructure in Schizosaccharomyces pombe using multi-color SMLM imaging.The Journal of cell biology · 2023Article
- Semi-automated 3D fluorescence speckle analyzer (3D-Speckler) for microscope calibration and nanoscale measurement.The Journal of cell biology · 2023Article
- Subcellular Euclidean distance measurements with multicolor fluorescence localization imaging in cultured cells.STAR protocols · 2021Article
- Trypanosome KKIP1 Dynamically Links the Inner Kinetochore to a Kinetoplastid Outer Kinetochore Complex.Frontiers in cellular and infection microbiology · 2021Article
- Effects of malleable kinetochore morphology on measurements of intrakinetochore tension.Open biology · 2020Article
- The Hec1/Ndc80 tail domain is required for force generation at kinetochores, but is dispensable for kinetochore-microtubule attachment formation and Ska complex recruitment.Molecular biology of the cell · 2020Article
- Ensemble-Level Organization of Human Kinetochores and Evidence for Distinct Tension and Attachment Sensors.Cell reports · 2020Article
- Aurora B kinase is recruited to multiple discrete kinetochore and centromere regions in human cells.The Journal of cell biology · 2020Article
- Mammalian kinetochores count attached microtubules in a sensitive and switch-like manner.The Journal of cell biology · 2019Article
- Complete microtubule-kinetochore occupancy favours the segregation of merotelic attachments.Nature communications · 2018Article
Corrections and comments
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Authors and funding
3 authors.
Funding
Abstract
Two-color fluorescence co-localization in 3D (three-dimension) has the potential to achieve accurate measurements at the nanometer length scale. Here, we optimized a 3D fluorescence co-localization method that uses mean values for chromatic aberration correction to yield the mean separation with ~10 nm accuracy between green and red fluorescently labeled protein epitopes within single human kinetochores. Accuracy depended critically on achieving small standard deviations in fluorescence centroid determination, chromatic aberration across the measurement field, and coverslip thickness. Computer simulations showed that large standard deviations in these parameters significantly increase 3D measurements from their true values. Our 3D results show that at metaphase, the protein linkage between CENP-A within the inner kinetochore and the microtubule-binding domain of the Ndc80 complex within the outer kinetochore is on average ~90 nm. The Ndc80 complex appears fully extended at metaphase and exhibits the same subunit structure in vivo as found in vitro by crystallography.
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