ArticleNature protocols2021
Single-particle tracking photoactivated localization microscopy of membrane proteins in living plant tissues.
Article in Nature protocols, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 33 papers.
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Who cites it
33 citing papers in PubMed.
- SRF3 couples cellular iron homeostasis and defense responses through ubiquitin-dependent nano-organization.Science advances · 2026Article
- Mutual antagonism between auxin and diacylglycerol acyltransferase 2 regulates high-temperature-induced hypocotyl elongation.The New phytologist · 2026Article
- Methods for the establishment of enzymatic mechanisms - from QM to ML.Chemical science · 2026Review
- The Arabidopsis SPFH protein HIR2 modulates receptor signaling and plasma membrane organization.The EMBO journal · 2026Article
- A practical guide to investigating biomolecular condensates: a comment from the plant community.Science China. Life sciences · 2026Review
- Mechanisms controlling the plasma membrane targeting and the nanodomain organization of the plant SPFH protein HIR2.The Plant journal : for cell and molecular biology · 2026Article
- Advanced Imaging for Live-Cell Spatiotemporal Monitoring: Technologies and Applications.Research (Washington, D.C.) · 2026Review
- Revealing plasma membrane protein dynamics in living plant cells with single-molecule tracking.Journal of experimental botany · 2025Article
- Technological advances in visualizing and rewiring microtubules during plant development.Journal of experimental botany · 2025Review
- Nanoscale spatiotemporal cluster analysis of expressed and endogenous proteins.Nature protocols · 2025Review
- Reciprocity in dynamics of supramolecular biosystems for the clustering of ligands and receptors.Proceedings of the National Academy of Sciences of the United States of America · 2025Article
- Broad application of the ALFA tagging technology for in planta nanobody-based imaging and biochemical characterization of plant proteins.Plant communications · 2025Article
- SlimVar for rapid in vivo single-molecule tracking of chromatin regulators in plants.Nature communications · 2025Article
- Modular in vivo assembly of Arabidopsis FCA oligomers into condensates competent for RNA 3' processing.The EMBO journal · 2025Article
- Single-molecule tracking in living microbial cells.Biophysics reports · 2025Article
- Trajectory Analysis in Single-Particle Tracking: From Mean Squared Displacement to Machine Learning Approaches.International journal of molecular sciences · 2024Review
- 4D Single-particle tracking with asynchronous read-out single-photon avalanche diode array detector.Nature communications · 2024Article
- Recent Advances in Fluorescent Nanoparticles for Stimulated Emission Depletion Imaging.Biosensors · 2024Review
- Diffusion and Oligomerization States of the Muscarinic MThe journal of physical chemistry. B · 2024Article
- Variable-Angle Epifluorescence Microscopy for Single-Particle Tracking in the Plant ER.Methods in molecular biology (Clifton, N.J.) · 2024Article
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Authors and funding
7 authors.
Funding
Abstract
Super-resolution microscopy techniques have pushed the limit of optical imaging to unprecedented spatial resolutions. However, one of the frontiers in nanoscopy is its application to intact living organisms. Here we describe the implementation and application of super-resolution single-particle tracking photoactivated localization microscopy (sptPALM) to probe single-molecule dynamics of membrane proteins in live roots of the model plant Arabidopsis thaliana. We first discuss the advantages and limitations of sptPALM for studying the diffusion properties of membrane proteins and compare this to fluorescence recovery after photobleaching (FRAP) and fluorescence correlation spectroscopy (FCS). We describe the technical details for handling and imaging the samples for sptPALM, with a particular emphasis on the specificity of imaging plant cells, such as their thick cell walls or high degree of autofluorescence. We then provide a practical guide from data collection to image analyses. In particular, we introduce our sptPALM_viewer software and describe how to install and use it for analyzing sptPALM experiments. Finally, we report an R statistical analysis pipeline to analyze and compare sptPALM experiments. Altogether, this protocol should enable plant researchers to perform sptPALM using a benchmarked reproducible protocol. Routinely, the procedure takes 3-4 h of imaging followed by 3-4 d of image processing and data analysis.
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Registered trials
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