Spatiotemporal multiplexed immunofluorescence imaging of living cells and tissues with bioorthogonal cycling of fluorescent probes.

Autor: Ko J; Center for Systems Biology, Massachusetts General Hospital, Boston, MA, USA., Wilkovitsch M; Institute of Applied Synthetic Chemistry, TU Wien, Vienna, Austria., Oh J; Center for Systems Biology, Massachusetts General Hospital, Boston, MA, USA., Kohler RH; Center for Systems Biology, Massachusetts General Hospital, Boston, MA, USA., Bolli E; Center for Systems Biology, Massachusetts General Hospital, Boston, MA, USA.; Department of Pathology and Immunology, University of Geneva, Geneva, Switzerland., Pittet MJ; Center for Systems Biology, Massachusetts General Hospital, Boston, MA, USA.; Department of Pathology and Immunology, University of Geneva, Geneva, Switzerland.; Ludwig Institute for Cancer Research, Lausanne Branch, Zurich, Switzerland.; AGORA Cancer Center, Lausanne, Switzerland., Vinegoni C; Center for Systems Biology, Massachusetts General Hospital, Boston, MA, USA., Sykes DB; Center for Regenerative Medicine, Massachusetts General Hospital, Boston, MA, USA.; Department of Medicine, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA., Mikula H; Institute of Applied Synthetic Chemistry, TU Wien, Vienna, Austria., Weissleder R; Center for Systems Biology, Massachusetts General Hospital, Boston, MA, USA. rweissleder@mgh.harvard.edu.; Department of Systems Biology, Harvard Medical School, Boston, MA, USA. rweissleder@mgh.harvard.edu., Carlson JCT; Center for Systems Biology, Massachusetts General Hospital, Boston, MA, USA. carlson.jonathan@mgh.harvard.edu.; Department of Medicine, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA. carlson.jonathan@mgh.harvard.edu.
Jazyk: angličtina
Zdroj: Nature biotechnology [Nat Biotechnol] 2022 Nov; Vol. 40 (11), pp. 1654-1662. Date of Electronic Publication: 2022 Jun 02.
DOI: 10.1038/s41587-022-01339-6
Abstrakt: Cells in complex organisms undergo frequent functional changes, but few methods allow comprehensive longitudinal profiling of living cells. Here we introduce scission-accelerated fluorophore exchange (SAFE), a method for multiplexed temporospatial imaging of living cells with immunofluorescence. SAFE uses a rapid bioorthogonal click chemistry to remove immunofluorescent signals from the surface of labeled cells, cycling the nanomolar-concentration reagents in seconds and enabling multiple rounds of staining of the same samples. It is non-toxic and functional in both dispersed cells and intact living tissues. We demonstrate multiparameter (n ≥ 14), non-disruptive imaging of murine peripheral blood mononuclear and bone marrow cells to profile cellular differentiation. We also show longitudinal multiplexed imaging of bone marrow progenitor cells as they develop into neutrophils over 6 days and real-time multiplexed cycling of living mouse hepatic tissues. We anticipate that SAFE will find broad utility for investigating physiologic dynamics in living systems.
(© 2022. The Author(s), under exclusive licence to Springer Nature America, Inc.)
Databáze: MEDLINE
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