{"doi":"10.1101/2023.07.11.548578","title":"Integrated Fluorescence Microscopy (iFLM) for Cryo-FIB-milling and In-situ Cryo-ET","abstract":"cryo-ET. However, correlative workflows that incorporate precise targeting remain challenging. Here, we demonstrate the development and use of an integrated Fluorescence Light Microscope (iFLM) module within a cryo-FIB-SEM to enable a coordinate-based two-point 3D correlative workflow. The iFLM guided targeting of regions of interest coupled with an automated milling process of the cryo-FIB-SEM instrument allows for the efficient preparation of 9-12 ∼200 nm thick lamellae within 24 hours. Using regular and montage-cryo-ET data collection schemes, we acquired data from FIB-milled lamellae of HeLa cells to examine cellular ultrastructure. Overall, this workflow facilitates on-the-fly targeting and automated FIB-milling of cryo-preserved cells, bacteria, and possibly high pressure frozen tissue, to produce lamellae for downstream cryo-ET data collection.","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2023,"id":390411,"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":11,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9502,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2023-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1162188,"name":"Veronika Vrbovská","orcid":"0000-0003-2863-6812","position":1,"is_corresponding":false},{"id":1162726,"name":"Tilman Franke","orcid":null,"position":2,"is_corresponding":false},{"id":575683,"name":"Bryan Sibert","orcid":"0000-0003-2650-2343","position":3,"is_corresponding":false},{"id":575682,"name":"Matt Larson","orcid":"0000-0003-2116-5747","position":4,"is_corresponding":false},{"id":1162189,"name":"Alex R. Hall","orcid":"0000-0002-3654-1373","position":5,"is_corresponding":false},{"id":1162190,"name":"Alexander Rigort","orcid":"0000-0002-4396-506X","position":6,"is_corresponding":false},{"id":1162727,"name":"John M. Mitchels","orcid":null,"position":7,"is_corresponding":false},{"id":420653,"name":"Elizabeth Wright","orcid":"0000-0003-0712-9475","position":8,"is_corresponding":false},{"id":414239,"name":"Jae Won Yang","orcid":"0000-0003-3689-5865","position":0,"is_corresponding":true}],"reference_count":28,"raw_metadata":null,"created_at":"2026-07-19T01:18:37.743462Z","pmid":"37502891","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":[]}