{"doi":"10.1117/1.nph.11.2.025005","title":"Two-dimensional electro-optical multiphoton microscopy","abstract":"SignificanceThe development of genetically encoded fluorescent indicators of neural activity with millisecond dynamics has generated demand for ever faster two-photon (2P) imaging systems, but acoustic and mechanical beam scanning technologies are approaching fundamental limits. We demonstrate that potassium tantalate niobate (KTN) electro-optical deflectors (EODs), which are not subject to the same fundamental limits, are capable of ultrafast two-dimensional (2D) 2P imaging in vivo.AimTo determine if KTN-EODs are suitable for 2P imaging, compatible with 2D scanning, and capable of ultrafast in vivo imaging of genetically encoded indicators with millisecond dynamics.ApproachThe performance of a commercially available KTN-EOD was characterized across a range of drive frequencies and laser parameters relevant to in vivo 2P microscopy. A second KTN-EOD was incorporated into a dual-axis scan module, and the system was validated by imaging signals in vivo from ASAP3, a genetically encoded voltage indicator.ResultsOptimal KTN-EOD deflection of laser light with a central wavelength of 960 nm was obtained up to the highest average powers and pulse intensities tested (power: 350 mW; pulse duration: 118 fs). Up to 32 resolvable spots per line at a 560 kHz line scan rate could be obtained with single-axis deflection. The complete dual-axis EO 2P microscope was capable of imaging a 13 μm by 13 μm field-of-view at over 10 kHz frame rate with ∼0.5 μm lateral resolution. We demonstrate in vivo imaging of neurons expressing ASAP3 with high temporal resolution.ConclusionsWe demonstrate the suitability of KTN-EODs for ultrafast 2P cellular imaging in vivo, providing a foundation for future high-performance microscopes to incorporate emerging advances in KTN-based scanning technology.","journal":"Neurophotonics","year":2024,"id":451507,"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":0.9568,"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":1273413,"name":"Samuel Stanek","orcid":null,"position":1,"is_corresponding":false},{"id":1091687,"name":"Harishankar Jayakumar","orcid":"0000-0002-1417-9860","position":2,"is_corresponding":false},{"id":292942,"name":"Zachary L. Newman","orcid":"0000-0002-4336-6585","position":3,"is_corresponding":false},{"id":915774,"name":"Jacob Gable","orcid":null,"position":4,"is_corresponding":false},{"id":1272940,"name":"James R. Leger","orcid":"0000-0001-9319-7196","position":5,"is_corresponding":false},{"id":1272941,"name":"Aaron Kerlin","orcid":"0000-0002-8257-5216","position":6,"is_corresponding":false},{"id":486663,"name":"Deano Farinella","orcid":"0000-0003-0312-0801","position":0,"is_corresponding":true}],"reference_count":41,"raw_metadata":null,"created_at":"2026-07-19T02:02:45.951080Z","pmid":"38841422","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":[]}