In Vivo Click Chemistry Enables Multiplexed Intravital Microscopy.

Adv Sci (Weinh)

Center for Systems Biology, Massachusetts General Hospital, 185 Cambridge St, CPZN 5206, Boston, MA, 02114, USA.

Published: August 2022

AI Article Synopsis

  • * This technique involves a bioorthogonal mechanism that can remove fluorescent signals from antibody-labeled cells quickly, enabling dynamic observation of specific cell types in real-time.
  • * The efficacy of this method is demonstrated through its application in live mice, allowing for complex imaging, including 12 color imaging of immune cells, while maintaining non-toxic conditions.

Article Abstract

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 (µm) 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.

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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9405492PMC
http://dx.doi.org/10.1002/advs.202200064DOI Listing

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