ArticleNature methods2024
ProDOL: a general method to determine the degree of labeling for staining optimization and molecular counting.
Article in Nature methods, 2024. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 6 papers.
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6 citing papers in PubMed.
- Characterising C-X-C Chemokine Receptor 4 Dynamics in the Cell Membrane Using Fluorescence Fluctuation Spectroscopy.Biomolecules · 2026Article
- Multivalent adaptor networks generate nanoscale organisation within T cell signalling condensates.bioRxiv : the preprint server for biology · 2026Article
- Super-Resolution Microscopy Reveals Nanoscale Arrangement of PD-L1 Immune Checkpoint.Chemical & biomedical imaging · 2026Article
- Quantitative Stimulated Emission Depletion (STED) Microscopy with DNA-Fluorophore Labels.ACS nano · 2026Article
- Single-molecule localization microscopy reveals the molecular organization of endogenous membrane receptors.Science advances · 2026Article
- Super-resolution microscopy for structural biology.Nature methods · 2025Review
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14 authors.
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Abstract
Determining the label to target ratio, also known as the degree of labeling (DOL), is crucial for quantitative fluorescence microscopy and a high DOL with minimal unspecific labeling is beneficial for fluorescence microscopy in general. Yet robust, versatile and easy-to-use tools for measuring cell-specific labeling efficiencies are not available. Here we present a DOL determination technique named protein-tag DOL (ProDOL), which enables fast quantification and optimization of protein-tag labeling. With ProDOL various factors affecting labeling efficiency, including substrate type, incubation time and concentration, as well as sample fixation and cell type can be easily assessed. We applied ProDOL to investigate how human immunodeficiency virus-1 pathogenesis factor Nef modulates CD4 T cell activation measuring total and activated copy numbers of the adapter protein SLP-76 in signaling microclusters. ProDOL proved to be a versatile and robust tool for labeling calibration, enabling determination of labeling efficiencies, optimization of strategies and quantification of protein stoichiometry.
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