Processing of fixed spheroids for TOBis barcoding, enzyme-free dissociation and antibody staining for CyTOF

This protocol is an adaptation and extension of the original work on single-cell signalling in organoids by the Tape Lab at UCL. In this adaptation, I have optimised a new approach for non-enzymatic single-cell dissociation of fixed, scaffold-free spheroids. The adapted protocol is also shorter and uses less material throughout.

Its development would not have been possible without the initial help provided by the Tape Lab in 2021-2022, particularly by Jahangir Sufi. For their original protocol, please refer to the following publication: Sufi, J., Qin, X., Rodriguez, F.C. et al. Multiplexed single-cell analysis of organoid signaling networks. Nat Protoc 16, 4897–4918 (2021). https://doi.org/10.1038/s41596-021-00603-4