Serial Immunoassays in Parallel on a Microfluidic Chip for Monitoring Hormone Secretion from Living Cells

• Published in: Analytical chemistry (Washington) Vol. 79; no. 3; pp. 947 - 954
• Main Authors: Dishinger, John F, Kennedy, Robert T
• Format: Journal Article
• Language: English
• Published: Washington, DC American Chemical Society 01.02.2007
• Subjects: Analytical chemistry; Cells - metabolism; Cellular biology; Chemistry; Chromatographic methods and physical methods associated with chromatography; Electrophoresis, Microchip - methods; Exact sciences and technology; Hormones; Hormones - analysis; Hormones - metabolism; Humans; Immunoassay; Immunoassay - instrumentation; Immunoassay - methods; Insulin - analysis; Insulin - metabolism; Insulin Secretion; Islets of Langerhans - metabolism; Microfluidic Analytical Techniques; Miscellaneous; Other chromatographic methods; Spectrometric and optical methods; Performance evaluation; Hormone; Antibody; Capillary electrophoresis; Laser induced fluorescence; Fluorescence spectrometry; Immunological method; Biological compound; Electroosmosis; Detection limit; Fluorescein; Isothiocyanates; Xanthene dye; Microfluidics; Monitoring
• ISSN: 0003-2700; 1520-6882
• DOI: 10.1021/ac061425s

A microfluidic chip that allows for the continuous monitoring of cellular secretions from multiple independent living samples was developed. Performance of the device was characterized through the analysis of insulin secretion from islets of Langerhans. The chip contained four individual channel networks, each capable of performing electrophoresis-based immunoassays of the perfusate from islets. In the networks, islets were housed in a chamber that was continuously perfused with pressure-driven biological media at 0.6 μL min-1. Electroosmosis was used to pull perfusate containing secreted insulin into 4-cm-long reaction channels where it mixed with fluorescein isothiocyanate-labeled insulin and anti-insulin antibody for 60 s. The reaction streams were sampled at 6.25-s intervals and analyzed in parallel using an on-chip capillary electrophoresis separation with laser-induced fluorescence detection by a scanning confocal microscope. The limit of detection for insulin was 10 nM. The device was used to complete over 1450 immunoassays of biological samples in less than 40 min, allowing the parallel monitoring of insulin release from four islets every 6.25 s.