Institute of Physiology, Faculty of Medicine, University of Maribor.
Institute of Physiology, Faculty of Medicine, University of Maribor; Faculty of Natural Sciences and Mathematics, University of Maribor.
J Vis Exp. 2021 Apr 13(170). doi: 10.3791/62293.
The acute mouse pancreatic tissue slice is a unique in situ preparation with preserved intercellular communication and tissue architecture that entails significantly fewer preparation-induced changes than isolated islets, acini, ducts, or dispersed cells described in typical in vitro studies. By combining the acute pancreatic tissue slice with live-cell calcium imaging in confocal laser scanning microscopy (CLSM), calcium signals can be studied in a large number of endocrine and exocrine cells simultaneously, with a single-cell or even subcellular resolution. The sensitivity permits the detection of changes and enables the study of intercellular waves and functional connectivity as well as the study of the dependence of physiological responses of cells on their localization within the islet and paracrine relationship with other cells. Finally, from the perspective of animal welfare, recording signals from a large number of cells at a time lowers the number of animals required in experiments, contributing to the 3R-replacement, reduction, and refinement-principle.
急性小鼠胰腺组织切片是一种独特的原位制备方法，具有保留的细胞间通讯和组织结构，与典型的体外研究中描述的分离胰岛、腺泡、导管或分散细胞相比，其引起的制备诱导变化要少得多。通过将急性胰腺组织切片与共聚焦激光扫描显微镜 (CLSM) 中的活细胞钙成像相结合，可以同时研究大量内分泌和外分泌细胞中的钙信号，具有单细胞甚至亚细胞分辨率。该方法的灵敏度允许检测变化，并能够研究细胞间波和功能连接，以及研究细胞的生理反应对其在胰岛内的定位和与其他细胞的旁分泌关系的依赖性。最后，从动物福利的角度来看，一次记录大量细胞的信号降低了实验所需动物的数量，有助于实现 3R 原则（替代、减少和优化）。
