title: Microfluidics platform for measurement of volume changes in immobilized intestinal enteroids.
abstract: Intestinal enteroids are ex vivo primary cultured single-layer epithelial cell spheroids of average diameter ∼150 μm with luminal surface facing inward. Measurement of enteroid swelling in response to secretagogues has been applied to genetic testing in cystic fibrosis and evaluation of drug candidates for cystic fibrosis and secretory diarrheas. The current measurement method involves manual addition of drugs and solutions to enteroids embedded in a Matrigel matrix and estimation of volume changes from confocal images of fluorescently stained enteroids. We developed a microfluidics platform for efficient trapping and immobilization of enteroids for quantitative measurement of volume changes. Multiple enteroids are trapped in a "pinball machine-like" array of polydimethylsiloxane posts for measurement of volume changes in unlabeled enteroids by imaging of an extracellular, high-molecular weight fluorescent dye. Measurement accuracy was validated using slowly expanding air bubbles. The method was applied to measure swelling of mouse jejunal enteroids in response to an osmotic challenge and cholera toxin-induced chloride secretion. The microfluidics platform allows for parallel measurement of volume changes on multiple enteroids during continuous superfusion, without an immobilizing matrix, and for quantitative volume determination without chemical labeling or assumptions about enteroid shape changes during swelling.
authors.affiliation: Departments of Medicine and Physiology, University of California, San Francisco, California 94143, USA.
authors.forename: Byung-Ju
authors.initials: BJ
authors.lastname: Jin
authors.affiliation: Departments of Medicine and Physiology, University of California, San Francisco, California 94143, USA.
authors.forename: Sailaja
authors.initials: S
authors.lastname: Battula
authors.affiliation: Departments of Physiology and Medicine, Gastroenterology Division, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA.
authors.forename: Nick
authors.initials: N
authors.lastname: Zachos
authors.affiliation: Departments of Physiology and Medicine, Gastroenterology Division, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA.
authors.forename: Olga
authors.initials: O
authors.lastname: Kovbasnjuk
authors.affiliation: Departments of Physiology and Medicine, Gastroenterology Division, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Fawlke-Abel
authors.affiliation: Departments of Physiology and Medicine, Gastroenterology Division, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA.
authors.forename: Julie
authors.initials: J
authors.lastname: In
authors.affiliation: Departments of Physiology and Medicine, Gastroenterology Division, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA.
authors.forename: Mark
authors.initials: M
authors.lastname: Donowitz
authors.affiliation: Departments of Medicine and Physiology, University of California, San Francisco, California 94143, USA.
authors.forename: Alan S
authors.initials: AS
authors.lastname: Verkman
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