{"doi":"10.1101/2025.06.02.657499","title":"OrganoidChip+: a Microfluidic Platform for Culturing, Staining, Immobilization, and High-Content Imaging of Adult Stem Cell-Derived Organoids","abstract":"High-content imaging (HCI) and analysis are the keys for advancing our understanding of the science behind organogenesis. To this end, culturing adult stem cell-derived organoids (ASOs) in a platform that also enables live imaging, staining, immobilization, and fast high-resolution imaging is crucial. However, existing platforms only partially satisfy these requirements. In this study, we present the OrganoidChip+, an all-in-one microfluidic device designed to integrate both culturing and HCI of ASOs all within one platform. We previously developed the OrganoidChip as a robust imaging tool. Now, the OrganoidChip+ incorporates several additional features for culturing organoids in addition to fluorescence staining and imaging without the need for sample transfer. The organoids grown within a culture chamber are stained and then transferred to immobilization chambers for blur-free, high-resolution imaging at predetermined locations. We cultured adult stem cell-derived intestinal organoids in the chip for 7 days and tracked growth rates of each organoid using intermittent brightfield images, followed by multiple image-based assays, including viability assay using widefield fluorescence imaging, a redox ratio assay using label-free, two-color, two-photon microscopy, and immunofluorescence assays using confocal microscopy. These assays serve as proof-of-concept to showcase the chip's capabilities in HCI of ASOs. Organoids cultured in the chip exhibited superior average growth rates over those in traditional Matrigel dome cultures, off-chip. Viability and redox ratio measurements of on-chip organoids were comparable or slightly better than their off-chip counterparts. Confocal imaging further confirmed that the OrganoidChip+ supports robust organoid culture while enabling detailed, high-resolution analysis. This all-in-one platform holds great potential for advancing ASO-based research, offering a scalable and cost-effective solution for HCI and analysis in organogenesis, drug screening, and disease modeling.","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2025,"id":555607,"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":0.9596,"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":1013056,"name":"Rishika Khanna","orcid":null,"position":1,"is_corresponding":false},{"id":1124060,"name":"Berk Camli","orcid":null,"position":2,"is_corresponding":false},{"id":1390222,"name":"Zeynep Nevra Yakay","orcid":null,"position":3,"is_corresponding":false},{"id":1072155,"name":"Anirudha Harihara","orcid":null,"position":4,"is_corresponding":false},{"id":1075005,"name":"Christopher Zdyrski","orcid":"0000-0002-7556-5298","position":5,"is_corresponding":false},{"id":561711,"name":"Sudip Mondal","orcid":"0000-0002-5387-0064","position":6,"is_corresponding":false},{"id":1072156,"name":"Evan Hegarty","orcid":null,"position":7,"is_corresponding":false},{"id":302786,"name":"Jonathan P. Mochel","orcid":"0000-0002-0997-3111","position":8,"is_corresponding":false},{"id":302785,"name":"Karin Allenspach","orcid":"0000-0002-2168-5809","position":9,"is_corresponding":false},{"id":382820,"name":"Adela Ben‐Yakar","orcid":"0000-0003-3468-6146","position":10,"is_corresponding":false},{"id":1071464,"name":"Khashayar Moshksayan","orcid":"0000-0002-5240-8193","position":0,"is_corresponding":true}],"reference_count":36,"raw_metadata":null,"created_at":"2026-07-19T02:54:59.329539Z","pmid":"40502189","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":[]}