ArticleJournal of experimental botany2011
Subcellular and single-molecule imaging of plant fluorescent proteins using total internal reflection fluorescence microscopy (TIRFM).
Article in Journal of experimental botany, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 25 papers.
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Who cites it
25 citing papers in PubMed.
- Clathrin-Mediated Endocytosis in Plants: Historical to Modern Advances.Traffic (Copenhagen, Denmark) · 2026Review
- Spatially resolved single-molecule profiling of microRNAs in migrating cells driven by microconfinement.Chemical science · 2022Article
- Article
- Quantifying the Organization and Dynamics of the Plant Plasma Membrane Across Scales Using Light Microscopy.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Correlative Light-Environmental Scanning Electron Microscopy of Plasma Membrane Efflux Carriers of Plant Hormone Auxin.Biomolecules · 2021Article
- Single-Molecule Imaging in Living Plant Cells: A Methodological Review.International journal of molecular sciences · 2021Review
- Current and future advances in fluorescence-based visualization of plant cell wall components and cell wall biosynthetic machineries.Biotechnology for biofuels · 2021Review
- Article
- Complementary Superresolution Visualization of Composite Plant Microtubule Organization and Dynamics.Frontiers in plant science · 2020Article
- Article
- Article
- Single Event Resolution of Plant Plasma Membrane Protein Endocytosis by TIRF Microscopy.Frontiers in plant science · 2017Article
- Real-time monitoring of NKCC2 endocytosis by total internal reflection fluorescence (TIRF) microscopy.American journal of physiology. Renal physiology · 2016Article
- Single-molecule fluorescence imaging to quantify membrane protein dynamics and oligomerization in living plant cells.Nature protocols · 2015Article
- Superresolution live imaging of plant cells using structured illumination microscopy.Nature protocols · 2015Article
- Dynamics and organization of cortical microtubules as revealed by superresolution structured illumination microscopy.Plant physiology · 2014Article
- Single-molecule detection and tracking in plants.Protoplasma · 2014Review
- A 'pocket guide' to total internal reflection fluorescence.Journal of microscopy · 2013Review
- Single cell optical imaging and spectroscopy.Chemical reviews · 2013Review
- AtFH1 formin mutation affects actin filament and microtubule dynamics in Arabidopsis thaliana.Journal of experimental botany · 2013Article
Corrections and comments
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Authors and funding
4 authors.
Funding
Abstract
Total internal reflection fluorescence microscopy (TIRFM) has been proven to be an extremely powerful technique in animal cell research for generating high contrast images and dynamic protein conformation information. However, there has long been a perception that TIRFM is not feasible in plant cells because the cell wall would restrict the penetration of the evanescent field and lead to scattering of illumination. By comparative analysis of epifluorescence and TIRF in root cells, it is demonstrated that TIRFM can generate high contrast images, superior to other approaches, from intact plant cells. It is also shown that TIRF imaging is possible not only at the plasma membrane level, but also in organelles, for example the nucleus, due to the presence of the central vacuole. Importantly, it is demonstrated for the first time that this is TIRF excitation, and not TIRF-like excitation described as variable-angle epifluorescence microscopy (VAEM), and it is shown how to distinguish the two techniques in practical microscopy. These TIRF images show the highest signal-to-background ratio, and it is demonstrated that they can be used for single-molecule microscopy. Rare protein events, which would otherwise be masked by the average molecular behaviour, can therefore be detected, including the conformations and oligomerization states of interacting proteins and signalling networks in vivo. The demonstration of the application of TIRFM and single-molecule analysis to plant cells therefore opens up a new range of possibilities for plant cell imaging.
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Registered trials
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