Optical Clearing and 3D Analysis Optimized for Mouse and Human Pancreata
Alexandra Alvarsson,
Maria Jimenez-Gonzalez,
Rosemary Li,
Carolina Rosselot,
Nikolaos Tzavaras,
Zhuhao Wu,
Sarah Stanley
Affiliations
Alexandra Alvarsson
Diabetes, Obesity, and Metabolism Institute, Division of Endocrinology, Diabetes and Bone Disease, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA
Maria Jimenez-Gonzalez
Diabetes, Obesity, and Metabolism Institute, Division of Endocrinology, Diabetes and Bone Disease, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA
Rosemary Li
Diabetes, Obesity, and Metabolism Institute, Division of Endocrinology, Diabetes and Bone Disease, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA
Carolina Rosselot
Diabetes, Obesity, and Metabolism Institute, Division of Endocrinology, Diabetes and Bone Disease, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA
Nikolaos Tzavaras
The Microscopy CoRE and Advanced Bioimaging Center, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA
Zhuhao Wu
Department of Cell, Developmental & Regenerative Biology, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USADepartment of Neuroscience, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA
Sarah Stanley
Diabetes, Obesity, and Metabolism Institute, Division of Endocrinology, Diabetes and Bone Disease, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USADepartment of Neuroscience, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA
The pancreas is a heavily innervated organ, but pancreatic innervation can be challenging to comprehensively assess using conventional histological methods. However, recent advances in whole-mount tissue clearing and 3D rendering techniques have allowed detailed reconstructions of pancreatic innervation. Optical clearing is used to enhance tissue transparency and reduce light scattering, thus eliminating the need to section the tissue. Here, we describe a modified version of the optical tissue clearing protocol iDISCO+ (immunolabeling-enabled three-dimensional imaging of solvent-cleared organs) optimized for pancreatic innervation and endocrine markers. The protocol takes 13-19 days, depending on tissue size. In addition, we include protocols for imaging using light sheet and confocal microscopes and for 3D segmentation of pancreatic innervation and endocrine cells using Imaris.
