Techniques to study nephron function: microscopy and imaging.

Recent advances in optics, computer sciences, fluorophores, and molecular techniques allow investigators the opportunity to study dynamic events within the functioning kidney with subcellular resolution. Investigators can now use two-photon microscopy to follow several complex heterogenous processes in organs such as the kidney with high spacial and temporal resolution. Repeat determinations over time within the same animal are possible and minimize animal use and interanimal variability. Furthermore, the ability to obtain volumetric data (3D) makes quantitative 4D (time) analysis possible. Finally, use of multiple fluorophores concurrently allows for three different or interactive processes to be observed simultaneously. Therefore, this approach compliments existing molecular, biochemical, and pharmacologic techniques by advancing in vivo data analysis and interpretation to subcellular levels for molecules without the requirement for fixation. Its use in the kidney is in its infancy but offers much promise for unraveling the complex interdependent physiologic and pathophysiologic processes known to contribute to cell function and disease.