{"doi":"10.1002/advs.202415648","title":"Gentle Label‐Free Nonlinear Optical Imaging Relaxes Linear‐Absorption‐Mediated Triplet","abstract":"Sample health is critical for live-cell fluorescence microscopy and has promoted light-sheet microscopy that restricts its ultraviolet-visible excitation to one plane inside a 3D sample. It is thus intriguing that laser-scanning nonlinear optical microscopy, which similarly restricts its near-infrared excitation, has not broadly enabled gentle label-free molecular imaging. It is hypothesized that intense near-infrared excitation induces phototoxicity via linear absorption of intrinsic biomolecules with subsequent triplet buildup, rather than the commonly assumed mechanism of nonlinear absorption. Using a reproducible phototoxicity assay based on the time-lapse elevation of autofluorescence (hyper-fluorescence) from a homogeneous tissue model (chicken breast), strong evidence is provided supporting this hypothesis. The study justifies a simple imaging technique, e.g., rapidly scanned sub-80-fs excitation with full triplet-relaxation, to mitigate this ubiquitous linear-absorption-mediated phototoxicity independent of sample types. The corresponding label-free imaging can track freely moving C. elegans in real-time at an irradiance up to one-half of water optical breakdown.","journal":"Advanced Science","year":2025,"id":544453,"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":1,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9636,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2025-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":530158,"name":"Lianhuang Li","orcid":"0000-0002-3531-6668","position":1,"is_corresponding":false},{"id":451761,"name":"Janet E. Sorrells","orcid":"0000-0003-0071-8509","position":2,"is_corresponding":false},{"id":530163,"name":"Jianxin Chen","orcid":"0000-0001-8519-4462","position":3,"is_corresponding":false},{"id":417936,"name":"Haohua Tu","orcid":"0000-0003-3116-0149","position":4,"is_corresponding":false},{"id":1167510,"name":"Geng Wang","orcid":"0000-0002-5615-4696","position":0,"is_corresponding":true}],"reference_count":53,"raw_metadata":null,"created_at":"2026-07-19T02:53:12.864581Z","pmid":"40432535","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":[]}