Measurement of intracellular calcium of submandibular glands using a high throughput plate reader

Calcium ions (Ca²⁺) impact nearly every aspect of cellular life and intracellular calcium [Ca²⁺]_i is a critical factor in the regulation of a plethora of physiological functions, including: muscle contraction, saliva secretion, metabolism, gene expression, cell survival and death. By measuring the changes of [Ca²⁺]_i levels, critical physiologic functions can be characterized and aberrant pathologic conditions or drug responses can be efficiently monitored. We developed a protocol for assessment of Ca²⁺ signaling in the acinar units of submandibular glands isolated from C57BL/6 mice, using benchtop, multi-mode, high throughput plate reader (FlexStation 3). This method represents a powerful tool for unlimited in vitro studies to monitor changes in receptor-mediated Ca²⁺ responses while retaining functional and morphological features of a native setting.