title: Microfluidic Chemical Cytometry for Enzyme Assays of Single Cells.
abstract: Cellular heterogeneity occurs, and should be probed, at multiple levels of cellular structure and physiology from the genome to enzyme activity. In particular, single-cell measures of protein levels are complemented by single-cell measurements of the activity of these proteins. Microfluidic assays of enzyme activity at the single-cell level combine moderate to high throughput with low dead volumes and the potential for automation. Herein, we describe the steps required to fabricate and operate a microfluidic device for chemical cytometry of fluorescent or fluorogenic reporters of enzyme activity in individual cells.
authors.affiliation: Department of Chemistry, Trinity College, 300 Summit Street, Hartford, CT, 06106, USA.
authors.forename: Livia
authors.initials: L
authors.lastname: Shehaj
authors.affiliation: Department of Chemistry, Trinity College, 300 Summit Street, Hartford, CT, 06106, USA.
authors.forename: Lorena
authors.initials: L
authors.lastname: Lazo de la Vega
authors.affiliation: Department of Chemistry, Trinity College, 300 Summit Street, Hartford, CT, 06106, USA. michelle.kovarik@trincoll.edu.
authors.forename: Michelle L
authors.initials: ML
authors.lastname: Kovarik
mesh_terms: D000818:Animals; D057075:Enzyme Assays / Q000295:instrumentation*; D004867:Equipment Design; D005336:Fiber Optic Technology; D005453:Fluorescence; D006801:Humans; D007834:Lasers; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D008856:Microscopy, Fluorescence; D055616:Microtechnology; D059010:Single-Cell Analysis / Q000295:instrumentation*
keywords: Chemical cytometry; Electrophoresis; Enzyme assay; Microchannel; Microfluidic; Reporter substrate