Multiplexed optical barcoding of cells via photochemical programming of bioorthogonal host–guest recognition
Open Access
- 19 February 2021
- journal article
- research article
- Published by Royal Society of Chemistry (RSC) in Chemical Science
- Vol. 12 (15), 5484-5494
- https://doi.org/10.1039/d0sc06860h
Abstract
Modern chemical and biological studies are undergoing a paradigm shift, where understanding the fate of individual cells, in an apparently homogeneous population, is becoming increasingly important. This has inculcated a growing demand for developing strategies that label individual cells with unique fluorescent signatures or barcodes so that their spatiotemporal trajectories can be mapped in real time. Among various approaches, light-regulated methods employing photocaged fluorophores have received particular attention, owing to their fine spatiotemporal control over labelling. However, their multiplexed use to barcode large numbers of cells for interrogating cellular libraries or complex tissues remains inherently challenging, due to the lack of multiple spectrally distinct photoactivated states in the currently available photocaged fluorophores. We report here an alternative multiplexable strategy based on optically controlled host–guest recognition in the cucurbit[7]uril (CB[7]) system that provides spatial control over the positioning of fluorophores to generate distinct barcodes in ‘user-defined’ cells. Using a combination of three spectrally distinct CB[7]-conjugated fluorophores and by sequentially performing cycles of photoactivation and fluorophore encoding, we demonstrate 10-color barcoding in microtubule-targeted fixed cells as well as 7-color barcoding in cell surface glycan targeted live MCF7 cells.Keywords
Funding Information
- Science and Engineering Research Board (CRG/2020/006183)
- The Wellcome Trust DBT India Alliance (IA/I/16/1/502368)
- Jawaharlal Nehru Centre for Advanced Scientific Research
- Department of Atomic Energy, Government of India (RTI 4007)
- Department of Science and Technology, Ministry of Science and Technology, India (CRG/2019/001219)
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