Protocol to measure insulin secretion dynamics from single mouse islets using a microfluidic fluorescence anisotropy immunoassay

The dynamics of glucose-stimulated insulin secretion (GSIS) from islets offer valuable insight into β cell metabolism. Here, we present a protocol for using a microfluidic device (i.e., InsC-chip) to measure the temporal dynamics of GSIS from individual mouse islets. We describe the steps for fabricating polydimethylsiloxane (PDMS)-based chips through soft lithography. We then present detailed procedures for employing the InsC-chip to measure insulin secretion from either four or eight individual islets, with the option for simultaneous live-cell imaging of the islet. For complete details on the use and execution of this protocol, please refer to Wang et al.<sup>1</sup>.

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Publication Details

Journal
STAR Protocols
Published
2025-11-20
DOI
https://doi.org/10.1016/j.xpro.2025.104209
Primary Topic
Pancreatic function and diabetes
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article
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article

Protocol to measure insulin secretion dynamics from single mouse islets using a microfluidic fluorescence anisotropy immunoassay

Jonathan V. Rocheleau, Yufeng Wang
STAR Protocols
Pancreatic function and diabetes
article

Protocol to measure insulin secretion dynamics from single mouse islets using a microfluidic fluorescence anisotropy immunoassay

Jonathan V. Rocheleau, Yufeng Wang
article en

Abstract

The dynamics of glucose-stimulated insulin secretion (GSIS) from islets offer valuable insight into β cell metabolism. Here, we present a protocol for using a microfluidic device (i.e., InsC-chip) to measure the temporal dynamics of GSIS from individual mouse islets. We describe the steps for fabricating polydimethylsiloxane (PDMS)-based chips through soft lithography. We then present detailed procedures for employing the InsC-chip to measure insulin secretion from either four or eight individual islets, with the option for simultaneous live-cell imaging of the islet. For complete details on the use and execution of this protocol, please refer to Wang et al.<sup>1</sup>.

STAR ProtocolsVol. 6(4)
University of Toronto (CA), Toronto General Hospital (CA)
Openalex Percentile: Top 46%
Pancreatic function and diabetes
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