Abstract
High-resolution three-dimensional imaging of fixed embryonic kidney tissues has advanced considerably in the past decade. Here we developed a new process for imaging whole metanephric organ culture at cell resolution in three dimensions over time. This technique combines the use of the newly available generation of infrared-optimized long working distance, high numerical aperture objectives and multiphoton fluorescence microscopy with a new system for vital staining of metanephric organ cultures with bodipy ceramide. This allows all cells in the organ culture to be visualized over time, enabling detailed observation of tissue morphogenesis. Thus, our method offers a powerful new approach for visualizing and understanding early events in renal development and for extending observations made in genetically manipulated models.