title: In situ, dual-mode monitoring of organ-on-a-chip with smartphone-based fluorescence microscope.
abstract: The use of organ-on-a-chip (OOC) platforms enables improved simulation of the human kidney's response to nephrotoxic drugs. The standard method of analyzing nephrotoxicity from existing OOC has majorly consisted of invasively collecting samples (cells, lysates, media, etc.) from an OOC. Such disruptive analyses potentiate contamination, disrupt the replicated in vivo environment, and require expertize to execute. Moreover, traditional analyses, including immunofluorescence microscopy, immunoblot, and microplate immunoassay are essentially not in situ and require substantial time, resources, and costs. In the present work, the incorporation of fluorescence nanoparticle immunocapture/immunoagglutination assay into an OOC enabled dual-mode monitoring of drug-induced nephrotoxicity in situ. A smartphone-based fluorescence microscope was fabricated as a handheld in situ monitoring device attached to an OOC. Both the presence of γ-glutamyl transpeptidase (GGT) on the apical brush-border membrane of 786-O proximal tubule cells within the OOC surface, and the release of GGT to the outflow of the OOC were evaluated with the fluorescence scatter detection of captured and immunoagglutinated anti-GGT conjugated nanoparticles. This dual-mode assay method provides a novel groundbreaking tool to enable the internal and external in situ monitoring of the OOC, which may be integrated into any existing OOCs to facilitate their subsequent analyses.
authors.affiliation: Department of Agricultural and Biosystems Engineering, The University of Arizona, Tucson, AZ, 85721-0038 USA.
authors.forename: Soohee
authors.initials: S
authors.lastname: Cho
authors.affiliation: Department of Pharmacology and Toxicology, The University of Arizona, Tucson, AZ, 85721-0038 USA.
authors.forename: Argel
authors.initials: A
authors.lastname: Islas-Robles
authors.affiliation: Biomedical Engineering Graduate Interdisciplinary Program, The University of Arizona, Tucson, AZ, 85721-0038 USA.
authors.forename: Ariana M
authors.initials: AM
authors.lastname: Nicolini
authors.affiliation: Department of Pharmacology and Toxicology, The University of Arizona, Tucson, AZ, 85721-0038 USA.
authors.forename: Terrence J
authors.initials: TJ
authors.lastname: Monks
authors.affiliation: Department of Agricultural and Biosystems Engineering, The University of Arizona, Tucson, AZ, 85721-0038 USA; Biomedical Engineering Graduate Interdisciplinary Program, The University of Arizona, Tucson, AZ, 85721-0038 USA. Electronic address: jyyoon@email.arizona.edu.
authors.forename: Jeong-Yeol
authors.initials: JY
authors.lastname: Yoon
mesh_terms: D000372:Agglutination Tests / Q000295:instrumentation; D001705:Bioprosthesis; D002460:Cell Line; D004867:Equipment Design; D019544:Equipment Failure Analysis; D006801:Humans; D007118:Immunoassay / Q000295:instrumentation* / Q000379:methods; D007668:Kidney / Q000187:drug effects / Q000276:immunology*; D007688:Kidneys, Artificial; D056656:Lab-On-A-Chip Devices; D008856:Microscopy, Fluorescence / Q000295:instrumentation* / Q000379:methods; D063731:Mobile Applications; D009924:Organ Culture Techniques / Q000295:instrumentation / Q000379:methods; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D000068997:Smartphone; D018675:Toxicity Tests / Q000295:instrumentation / Q000379:methods; D014584:User-Computer Interface
keywords: 786-O cells; Kidney-on-a-chip; Particle immunoagglutination; Smartphone microscope; γ-glutamyl transpeptidase