Takayama Jun, Fujita Masashi, Onami Shuichi
RIKEN Quantitative Biology Center, Laboratory for Developmental Dynamics, Kobe, Japan.
Bio Protoc. 2017 Apr 5;7(7):e2205. doi: 10.21769/BioProtoc.2205.
Fertilization calcium waves are a conserved trigger for animal development; however, genetic analysis of these waves has been limited due to the difficulty of imaging fertilization. Here we describe a protocol to image calcium dynamics during fertilization in the genetic animal model . This protocol consists of germline microinjection of a chemical calcium indicator, worm immobilization, live imaging, and image processing that quantifies the calcium fluorescence in the oocyte region moving in the field-of-view during ovulation. This imaging protocol can also be used to image other cellular processes during fertilization in , such as membrane fusion and cytoskeletal dynamics.
受精钙波是动物发育中一种保守的触发因素；然而，由于受精成像困难，对这些钙波的遗传分析一直有限。在这里，我们描述了一种在遗传动物模型中对受精过程中的钙动力学进行成像的方案。该方案包括对化学钙指示剂进行种系显微注射、使蠕虫固定、实时成像以及图像处理，该图像处理可量化排卵期间在视野中移动的卵母细胞区域的钙荧光。这种成像方案还可用于对受精过程中的其他细胞过程进行成像，如膜融合和细胞骨架动力学。
