Kim Abraham, Kilimnik German, Hara Manami
Department of Medicine, University of Chicago, USA.
J Vis Exp. 2010 Jun 7(40):1970. doi: 10.3791/1970.
Tracing changes of specific cell populations in health and disease is an important goal of biomedical research. The process of monitoring pancreatic beta-cell proliferation and islet growth is particularly challenging. We have developed a method to capture the distribution of beta-cells in the intact pancreas of transgenic mice with fluorescence-tagged beta-cells with a macro written for ImageJ (rsb.info.nih.gov/ij/). Following pancreatic dissection and tissue clearing, the entire pancreas is captured as a virtual slice, after which the GFP-tagged beta-cells are examined. The analysis includes the quantification of total beta-cell area, islet number and size distribution with reference to specific parameters and locations for each islet and for small clusters of beta-cells. The entire distribution of islets can be plotted in three dimensions, and the information from the distribution on the size and shape of each islet allows a quantitative and qualitative comparison of changes in overall beta-cell area at a glance.
追踪健康和疾病状态下特定细胞群的变化是生物医学研究的一个重要目标。监测胰腺β细胞增殖和胰岛生长的过程尤其具有挑战性。我们开发了一种方法，使用为ImageJ（rsb.info.nih.gov/ij/）编写的宏，来捕获荧光标记β细胞的转基因小鼠完整胰腺中β细胞的分布。在胰腺解剖和组织透明化之后，整个胰腺被捕获为一个虚拟切片，然后检查绿色荧光蛋白标记的β细胞。该分析包括参考每个胰岛和小β细胞簇的特定参数及位置，对总β细胞面积、胰岛数量和大小分布进行量化。胰岛的整个分布可以三维绘制，每个胰岛大小和形状分布的信息使得能够一眼对总β细胞面积的变化进行定量和定性比较。
