A1 Refereed original research article in a scientific journal
A specific hybridisation internalisation probe (SHIP) enables precise live-cell and super-resolution imaging of internalized cargo
Authors: Hernández-Pérez Sara, Mattila Pieta K.
Publisher: NATURE PORTFOLIO
Publication year: 2022
Journal: Scientific Reports
Journal name in source: SCIENTIFIC REPORTS
Journal acronym: SCI REP-UK
Article number: 620
Volume: 12
Issue: 1
Number of pages: 15
ISSN: 2045-2322
DOI: https://doi.org/10.1038/s41598-021-04544-6
Web address : https://www.nature.com/articles/s41598-021-04544-6
Self-archived copy’s web address: https://research.utu.fi/converis/portal/detail/Publication/174760351
Facilitated by the advancements in microscopy, our understanding of the complexity of intracellular vesicle traffic has dramatically increased in recent years. However, distinguishing between plasma membrane-bound or internalised ligands remains a major challenge for the studies of cargo sorting to endosomal compartments, especially in small and round cells such as lymphocytes. The specific hybridization internalisation probe (SHIP) assay, developed for flow cytometry studies, employs a ssDNA fluorescence internalisation probe and a complementary ssDNA quenching probe to unambiguously detect the internalized receptors/cargo. Here, we adopted the SHIP assay to study the trafficking of receptor/ligand complexes using B lymphocytes and B cell receptor-mediated antigen internalization as a model system. Our study demonstrates the potential of the SHIP assay for improving the imaging of internalized receptor/ligand complexes and establishes the compatibility of this assay with multiple imaging modalities, including live-cell imaging and super-resolution microscopy.
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