{"doi":"10.1101/2024.03.08.584185","title":"Click-free imaging of carbohydrate trafficking in live cells using an azido photothermal probe","abstract":"Real-time tracking of intracellular carbohydrates remains challenging. While click chemistry allows bio-orthogonal tagging with fluorescent probes, the reaction permanently alters the target molecule and only allows a single snapshot. Here, we demonstrate click-free mid-infrared photothermal (MIP) imaging of azide-tagged carbohydrates in live cells. Leveraging the micromolar detection sensitivity for 6-azido-trehalose (TreAz) and the 300-nm spatial resolution of MIP imaging, the trehalose recycling pathway in single mycobacteria, from cytoplasmic uptake to membrane localization, is directly visualized. A peak shift of azide in MIP spectrum further uncovers interactions between TreAz and intracellular protein. MIP mapping of unreacted azide after click reaction reveals click chemistry heterogeneity within a bacterium. Broader applications of azido photothermal probes to visualize the initial steps of the Leloir pathway in yeasts and the newly synthesized glycans in mammalian cells are demonstrated.","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2024,"id":486798,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":3,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9571,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2024-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1330568,"name":"Harini A. Perera","orcid":"0000-0002-9514-6170","position":1,"is_corresponding":false},{"id":1330569,"name":"Rylie Bolarinho","orcid":"0000-0002-8620-3583","position":2,"is_corresponding":false},{"id":1330570,"name":"Zeke A. Piskulich","orcid":"0000-0003-0304-305X","position":3,"is_corresponding":false},{"id":468327,"name":"Zhongyue Guo","orcid":"0009-0008-0780-186X","position":4,"is_corresponding":false},{"id":629770,"name":"Jiaze Yin","orcid":"0000-0001-6080-3073","position":5,"is_corresponding":false},{"id":291111,"name":"Hongjian He","orcid":"0000-0003-3030-8279","position":6,"is_corresponding":false},{"id":1145258,"name":"Mingsheng Li","orcid":"0000-0002-7977-0854","position":7,"is_corresponding":false},{"id":817222,"name":"Xiaowei Ge","orcid":"0000-0001-8938-9304","position":8,"is_corresponding":false},{"id":267953,"name":"Qiang Cui","orcid":"0000-0001-6214-5211","position":9,"is_corresponding":false},{"id":675718,"name":"Olof Ramström","orcid":"0000-0002-1533-6514","position":10,"is_corresponding":false},{"id":463185,"name":"Mingdi Yan","orcid":"0000-0003-1121-4007","position":11,"is_corresponding":false},{"id":266496,"name":"Ji‐Xin Cheng","orcid":"0000-0002-5607-6683","position":12,"is_corresponding":false},{"id":1047628,"name":"Qing Xia","orcid":"0000-0002-5939-2972","position":0,"is_corresponding":true}],"reference_count":53,"raw_metadata":null,"created_at":"2026-07-19T02:08:01.404471Z","pmid":"38559219","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}