Quantitative monitoring of insulin secretion from single islets of Langerhans in parallel on a microfluidic chip.
Quantification of insulin release from pancreatic islets of Langerhans is of interest for diabetes research. Typical insulin secretion experiments are performed using offline techniques that are expensive, slow, have low-throughput, and require multiple islets. We have developed a microfluidic device for high-throughput, automated, and online monitoring of insulin secretion from individual islets in parallel. This chip consists of 15 channel networks each capable of superfusing a single islet and mixing superfusate from each islet online with fluorescein isothiocyanate-labeled insulin and anti-insulin antibody for a competitive immunoassay. The resulting continuous reaction streams are periodically injected onto parallel electrophoresis channels where the mixtures are separated. The resulting traces are used to quantify relative insulin released from islets. Serial immunoassays were performed at 10 s intervals on all 15 channels, corresponding to 5400 immunoassays per hour, to create temporally resolved insulin release profiles that captured single islet secretion dynamics. The chip was used to demonstrate that free fatty acid induced lipotoxicity in islets eliminates pulsatile insulin secretion.
Dishinger
John F
JF
Department of Chemistry, University of Michigan, Ann Arbor, Michigan 48109, USA.
Reid
Kendra R
KR
Kennedy
Robert T
RT
D000818:Animals; D002417:Cattle; D019075:Electrophoresis, Capillary; D005227:Fatty Acids / Q000494:pharmacology; D005453:Fluorescence; D007328:Insulin / Q000378:metabolism*; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D007801:Langerhans Cells / Q000187:drug effects / Q000378:metabolism*; D046210:Microfluidic Analytical Techniques; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013997:Time Factors
D005227:Fatty Acids; D007328:Insulin