University of Nevada Reno School of Medicine

Genetically encoded Ca²⁺ indicators (GECIs) have radically changed how in situ Ca²⁺ imaging is performed. To maximize data recovery from such recordings, appropriate analysis of Ca²⁺ signals is required. The protocols in this paper facilitate the quantification of Ca²⁺ signals recorded in situ using spatiotemporal mapping and particle-based analysis.

The detrusor-free bladder model enables direct access to the suburothelium to study local mechanisms for regulation of biologically active mediator availability in suburothelium/lamina propria during storage and voiding of urine. The preparation closely resembles filling of an intact bladder and allows pressure-volume studies to be performed without systemic influences.

The goal of this protocol is to isolate intact pacemaker regions of the mouse renal pelvis using vibratome sectioning. These sections can then be used for in situ Ca²⁺ imaging to elucidate Ca²⁺ transient properties of pacemaker cells and other interstitial cells in vibratome slices.