The mineralocorticoid aldosterone is produced by zona glomerulosa cells of the adrenal cortex upon stimulation by several secretagogues. The most relevant ones, angiotensin II and hyperkalemia, trigger aldosterone production by increasing cytoplasmic Ca²⁺ activity. Up to now, primary cultured cells or cell lines derived from the adrenal cortex have been the mostly used models for the studying the signaling pathways involved in aldosterone production. However, many pitfalls like cell dedifferentiation or the difficult identification of a particular cell type in a mixed primary culture remained insurmountable until now. To overcome these problems we adapted the slice technique for adrenocortical tissue. Freshly prepared murine adrenal slices were loaded with the Ca²⁺ sensitive dye Fluo-4 and the Ca²⁺ responses of zona glomerulosa cells upon angiotensin II and high extracellular K⁺ concentrations were determined. Compared to measurements in primary cells, measurements in slices are accomplished in a more physiological scenario, where cell interactions and organ structure are largely preserved. This technique allowed to determine the cellular sub-type (glomerulosa vs fasciculata cells) and gave valuable insights in the Ca²⁺ signaling of adrenocortical cells. In the future we wish to adapt this method for human tissue samples. Such an approach can shed light on important mechanisms underlying pathological adrenal hormone secretion in human diseases.