Nanoliter Cell Culture Array with Tunable Chemical Gradients

Analytical Chemistry
Jonathan AvesarShulamit Levenberg

Abstract

A multitude of cell screening assays for diagnostic and research applications rely on quantitative measurements of a sample in the presence of different reagent concentrations. Standard methods rely on microtiter plates of varying well density, which provide simple and standardized sample addressability. However, testing hundreds of chemical dilutions requires complex automation, and typical well volumes of microtiter plates are incompatible with the analysis of a small number of cells. Here, we present a microfluidic device for creating a high-resolution chemical gradient spanning 200 nanoliter wells. Using air-based shearing, we show that the individual wells can be compartmentalized without altering the concentration gradient, resulting in a large set of isolated nanoliter cell culture wells. We provide an analytical and numerical model for predicting the concentration within each culture chamber and validate it against experimental results. We apply our system for the investigation of yeast cell metabolic gene regulation in the presence of different ratios of galactose/glucose concentrations and successfully resolve the nutrient threshold at which the cells activate the galactose pathway.

References

Jun 1, 2007·Lab on a Chip·Elisa FigalloGordana Vunjak-Novakovic
May 4, 2010·Trends in Biotechnology·Daojing Wang, Steven Bodovitz
Feb 10, 2012·Lab on a Chip·Mira T GuoDavid A Weitz
Mar 30, 2012·Lab on a Chip·Jeongyun KimArul Jayaraman
Sep 12, 2012·Genetics·James R Broach
Nov 28, 2012·Angewandte Chemie·Haakan N Joensson, Helene Andersson Svahn
Feb 19, 2013·BMC Molecular Biology·Christopher J WeltyColm Morrissey
Mar 29, 2014·Lab on a Chip·Jonathan AvesarShulamit Levenberg
Jul 24, 2014·Proceedings of the National Academy of Sciences of the United States of America·Jonathan ShemeshShulamit Levenberg
Jan 22, 2015·Proceedings of the National Academy of Sciences of the United States of America·Renan Escalante-ChongMichael Springer
Oct 22, 2015·Biomicrofluidics·Cyprien GuermonprezCharles N Baroud
Jun 28, 2017·Proceedings of the National Academy of Sciences of the United States of America·Jonathan AvesarShulamit Levenberg

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Citations

Feb 14, 2021·Cancers·Fahriye DuzagacFlavio Rizzolio
Apr 1, 2021·Analytical Chemistry·Ann V NguyenAlireza Abbaspourrad
Nov 11, 2020·Analytical Chemistry·Simon F BerlandaPetra S Dittrich

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