TY  - JOUR
TI  - Preparation of Whole-mount Mouse Islets on Vascular Extracellular Matrix for Live Islet Cell Microscopy
AU  - Ho, Kung- Hsien
AU  - Gu, Guoqiang 
AU  - Kaverina, Irina 
VL  - 13
IS  - 21
PY  - 2023
DA  - 2023/11/05
SP  - e4868
C1  - Bio-protocol 2023;13:e4868
DO  - 10.21769/BioProtoc.4868
UR  - https://doi.org/10.21769/BioProtoc.4868
AB  - Pancreatic islet β cells preferentially secrete insulin toward the plasma membrane, making contact with the capillary extracellular matrix (ECM). Isolated islets separated from the exocrine acinar cells are the best system for cell biology studies of primary β cells, whereas isolated islets lose their capillary network during ex vivo culture. Providing the appropriate extracellular signaling by attaching islets to vascular ECM-coated surfaces can restore the polarized insulin secretion toward the ECM. The guided secretion toward ECM-coated glass coverslips provides a good model for recording insulin secretion in real time to study its regulation. Additionally, β cells attached to the ECM-coated coverslips are suitable for confocal live imaging of subcellular components including adhesion molecules, cytoskeleton, and ion channels. This procedure is also compatible for total internal reflection fluorescence (TIRF) microscopy, which provides optimal signal-to-noise ratio and high spatial precision of structures close to the plasma membrane. In this article, we describe the optimized protocol for vascular ECM-coating of glass coverslips and the process of attachment of isolated mouse islets on the coverslip. This preparation is compatible with any high-resolution microscopy of live primary β cells.Key features• Optimized coating procedure to attach isolated islets, compatible for both confocal and TIRF microscopy.• The ECM-coated glass coverslip functions as the artificial capillary surface to guide secretion toward the coated surface for optimal imaging of secretion events.• Shows the process of islets attachment to the ECM-coated surface in a 6-day ex vivo culture.Graphical overview
KW  - Pancreatic β cells
KW  - Islet attachment
KW  - Islet ex vivo culture
KW  - Islet flattening
KW  - Human ECM
KW  - Live islet cells visualization
JF  - Bio-protocol
SN  - 2331-8325
PB  - Bio-protocol LLC.
BIO101  - False