Refereed journal article or data article (A1)

A specific hybridisation internalisation probe (SHIP) enables precise live-cell and super-resolution imaging of internalized cargo




List of 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

Volume number: 12

Issue number: 1

Number of pages: 15

ISSN: 2045-2322

DOI: http://dx.doi.org/10.1038/s41598-021-04544-6

URL: https://www.nature.com/articles/s41598-021-04544-6

Self-archived copy’s web address: https://research.utu.fi/converis/portal/detail/Publication/174760351


Abstract
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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Last updated on 2022-28-06 at 14:38