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A Multi-Parametric Islet Perifusion System within a Microfluidic Perifusion Device
Published on: January 26, 2010
Protocol to measure insulin secretion dynamics from single mouse islets using a microfluidic fluorescence anisotropy
Yufeng Wang1, Jonathan V Rocheleau2
1Advanced Diagnostics, Toronto General Hospital Research Institute, Toronto, ON M5G 1L7, Canada; Institute of Biomedical Engineering, University of Toronto, Toronto, ON M5S 3G9, Canada.
STAR Protocols
|November 19, 2025
Summary
This study details a new microfluidic InsC-chip protocol for measuring glucose-stimulated insulin secretion (GSIS) dynamics in individual mouse islets. The method allows for precise temporal analysis of beta cell function using live-cell imaging.
Area of Science:
- Endocrinology
- Cell Biology
- Biomedical Engineering
Background:
- Glucose-stimulated insulin secretion (GSIS) is crucial for maintaining glucose homeostasis.
- Understanding beta cell metabolism dynamics is key to addressing metabolic diseases.
- Existing methods for measuring GSIS from individual islets have limitations.
Purpose of the Study:
- To present a detailed protocol for fabricating and utilizing a microfluidic InsC-chip.
- To enable precise measurement of temporal GSIS dynamics from individual mouse islets.
- To facilitate simultaneous live-cell imaging during insulin secretion measurements.
Main Methods:
- Fabrication of polydimethylsiloxane (PDMS)-based microfluidic chips via soft lithography.
- Detailed procedures for loading and perfusing individual mouse islets (4 or 8 per chip).
- Integration of live-cell imaging capabilities for real-time observation of islet responses.
Main Results:
- Successful demonstration of the InsC-chip protocol for measuring temporal GSIS.
- High-resolution data on insulin secretion patterns from single islets.
- Validation of the chip's suitability for simultaneous functional and imaging studies.
Conclusions:
- The InsC-chip provides a robust platform for studying beta cell function at the single-islet level.
- This protocol advances the ability to investigate the temporal dynamics of GSIS.
- The InsC-chip is a valuable tool for research in diabetes and metabolic disorders.
