Open AccessIn Vivo Click Chemistry Enables Multiplexed Intravital Microscopy
Author(s) -
Ko Jina,
Lucas Kilean,
Kohler Rainer,
Halabi Elias A.,
Wilkovitsch Martin,
Carlson Jonathan C. T.,
Weissleder Ralph
Publication year - 2022
Publication title -
advanced science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.388
H-Index - 100
ISSN - 2198-3844
DOI - 10.1002/advs.202200064
Subject(s) - in vivo , bioorthogonal chemistry , preclinical imaging , click chemistry , intravital microscopy , fluorophore , chemistry , biophysics , fluorescence microscope , ex vivo , microbiology and biotechnology , biology , fluorescence , combinatorial chemistry , optics , physics
The ability to observe cells in live organisms is essential for understanding their function in complex in vivo milieus. A major challenge today has been the limited ability to perform higher multiplexing beyond four to six colors to define cell subtypes in vivo. Here, a click chemistry‐based strategy is presented for higher multiplexed in vivo imaging in mouse models. The method uses a scission‐accelerated fluorophore exchange (SAFE), which exploits a highly efficient bioorthogonal mechanism to completely remove fluorescent signal from antibody‐labeled cells in vivo. It is shown that the SAFE‐intravital microscopy imaging method allows 1) in vivo staining of specific cell types in dorsal and cranial window chambers of mice, 2) complete un‐staining in minutes, 3) in vivo click chemistries at lower (µ m ) and thus non‐toxic concentrations, and 4) the ability to perform in vivo cyclic imaging. The potential utility of the method is demonstrated by 12 color imaging of immune cells in live mice.