Spatiotemporal dynamics and functions of intracellular Ca²⁺ signaling in peripheral organs, such as pancreatic β-cells and liver hepatocytes, have been studied using in vitro and ex vivo preparations. These preparations are free from intravital environment including the influence of the autonomic nervous system, hormones, other bioactive substances and cell-to-cell interactions. Thus, Ca²⁺ dynamics in these cells under physiological conditions have not been clarified. In vivo Ca²⁺ imaging analysis of these cells remains challenging, therefore, we are trying to establish it with transgenic mouse lines expressing Ca²⁺ indicator proteins. We here report the findings and perspectives using our method.