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Updated: Sep 21, 2025
Homogeneous Time-resolved Förster Resonance Energy Transfer-based Assay for Detection of Insulin Secretion
Published on: May 10, 2018
Increasing insulin measurement throughput by fluorescence anisotropy imaging immunoassays
Yao Wang1, Damilola I Adeoye1, Yue J Wang2
1Department of Chemistry and Biochemistry, Florida State University, 95 Chieftain Way, Tallahassee, FL, 32306, USA.
Insights
Researchers developed a new microfluidic system to measure insulin secretion from single islets of Langerhans. This fluorescence anisotropy imaging immunoassay offers higher throughput for studying dynamic glucose-regulating hormone release.
Area of Science:
- Endocrinology
- Biomedical Engineering
- Analytical Chemistry
Background:
- Insulin, secreted by islets of Langerhans, is crucial for blood glucose regulation.
- Single islet analysis reveals heterogeneous insulin secretion patterns, necessitating advanced measurement techniques.
- Existing methods lack the throughput to analyze dynamic release from small islet groups in parallel.
Purpose of the Study:
- To develop and validate a microfluidic system for high-throughput, parallel measurement of insulin secretion from islets of Langerhans.
- To assess the feasibility of fluorescence anisotropy imaging immunoassays for dynamic islet secretion studies.
- To investigate functional heterogeneity in insulin release at the single islet level.
Main Methods:
- A microfluidic device utilizing vacuum pressure to draw perfusate and reagents through 12 parallel mixing channels for competitive immunoassays.
- Fluorescence anisotropy imaging employing polarized light excitation and emission detection with a sCMOS camera.
- Parallel monitoring of insulin release from groups of murine and human islets.
Main Results:
- The system demonstrated the capability to measure dynamic insulin release from multiple islet chambers simultaneously.
- Observed heterogeneity in insulin secretion traces from individual islets.
- Identified challenges in precise insulin quantification due to islet-induced variations in chamber resistance.
Conclusions:
- The developed microfluidic fluorescence anisotropy imaging immunoassay is a promising advancement for increasing the throughput of islet secretion measurements.
- Despite quantification challenges, the system provides valuable insights into islet functional heterogeneity.
- Further optimization is needed to overcome resistance-related issues for more accurate hormone quantification.
Abstract:
Insulin secreted from islets of Langerhans is the main hormone to reduce blood glucose. Examination of insulin secretion patterns at the single islet level reveals functional differences in the timings and patterns of release. This heterogeneous response highlights the importance of developing systems to measure dynamic release from small numbers of islets in parallel. Toward this, we describe fluorescence anisotropy imaging immunoassays as a relatively simple method for increased throughput of islet secretion measurements. In this system, vacuum pressure from a syringe pump pulled perfusate from 12 islet chambers and reagents into 12 parallel mixing channels for a competitive immunoassay. Light from a Xe arc lamp was filtered and polarized prior to focusing on the microfluidic device at the region where the 12 mixing channels converged. Emission was collected and passed through vertical and horizontal emission polarizers housed in an automated filter wheel before being imaged with a sCMOS camera for the determination of anisotropy. This microfluidic system was tested by monitoring insulin release from groups of murine and human islets. Heterogeneity was observed in the islet traces; however, the presence of islets affected the resistance of the islet chambers, hampering insulin quantification. Nonetheless, this microfluidic system is a step towards increasing the throughput of hormone release measurements from islets of Langerhans.
