ArticleMethods in molecular biology (Clifton, N.J.)2024
Variable-Angle Epifluorescence Microscopy for Single-Particle Tracking in the Plant ER.
Article in Methods in molecular biology (Clifton, N.J.), 2024. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 4 papers.
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Who cites it
4 citing papers in PubMed, 4 citations in OpenAlex.
- Built different: ER cisternae formed by the Arabidopsis Lunapark proteins differ in ultrastructure and affect ER-Golgi transport.The New phytologist · 2026Article
- Revealing plasma membrane protein dynamics in living plant cells with single-molecule tracking.Journal of experimental botany · 2025Article
- SlimVar for rapid in vivo single-molecule tracking of chromatin regulators in plants.Nature communications · 2025Article
- OneFlowTraX: a user-friendly software for super-resolution analysis of single-molecule dynamics and nanoscale organization.Frontiers in plant science · 2024Article
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Authors and funding
4 authors at 3 institutions in 1 country.
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Abstract
Single-particle tracking (SPT) of biomolecules in the plant endoplasmic reticulum has the potential to inform on the formation of protein-protein complexes, metabolons, and the transport of molecules through both the ER membrane and lumen. Plant cells are particularly challenging for observing and tracking single molecules due to their unique structure, size, and considerable autofluorescence. However, by using variable-angle or highly inclined epifluorescence microscopy (VAEM) and transient expression in tobacco, it is possible to observe single-particle dynamics in the ER. Selecting the appropriate fluorophore, and ensuring the correct fluorophore density in the ER, is essential for successful SPT. By using tuneable fluorophores, which can be photoconverted and photoactivated, it is possible to vary the density of visible fluorophores in the ER dynamically. Here we describe methods to prepare plant samples for VAEM and two methods for determining and analyzing single-particle tracks from VAEM time series.
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Registered trials
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