Optical Clearing and 3D Analysis Optimized for Mouse and Human Pancreata.
Optical Clearing and 3D Analysis Optimized for Mouse and Human Pancreata.
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DOI：
10.21769/bioprotoc.4103
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发表时间：
2021-08
期刊：
影响因子：
0.8
通讯作者：
A. Alvarsson;M. Jimenez-Gonzalez;Rosemary Li;Carolina Rosselot;N. Tzavaras;Zhuhao Wu;S. Stanley
中科院分区：
文献类型：
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作者：
A. Alvarsson;M. Jimenez-Gonzalez;Rosemary Li;Carolina Rosselot;N. Tzavaras;Zhuhao Wu;S. Stanley
胰腺是一个高度神经支配的器官，但胰腺神经支配可能具有挑战性，以全面评估使用传统的组织学方法。然而，最近的进展，在整体组织清除和3D渲染技术允许胰腺神经支配的详细重建。光学透明用于增强组织透明度并减少光散射，从而消除对组织切片的需要。在这里，我们描述了一个修改后的版本的光学组织清除协议iDISCO+（免疫标记使能的三维成像的溶剂清除器官）优化胰腺神经支配和内分泌标志物。该方案需要13-19天，取决于组织大小。此外，我们还包括使用光片和共聚焦显微镜成像的协议，以及使用Imaris对胰腺神经支配和内分泌细胞进行3D分割的协议。
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.