Using 2-photon microscopy to understand albuminuria.

Intravital 2-photon microscopy, along with the development of fluorescent probes and innovative software, has rapidly advanced the study of intracellular and intercellular processes at the organ level. Researchers can quantify the distribution, behavior, and dynamic interactions of up to four labeled chemical probes and proteins simultaneously and repeatedly in four dimensions (3D + time) with subcellular resolution in real time. Transgenic fluorescently labeled proteins, delivery of plasmids, and photo-activatable probes enhance these possibilities. Thus, multi-photon microscopy has greatly extended our ability to understand cell biology intra-vitally at cellular and subcellular levels. For example, evaluation of rat surface glomeruli and accompanying proximal tubules has shown the long held paradigm regarding limited albumin filtration under physiologic conditions is to be questioned. Furthermore, the role of proximal tubules in determining albuminuria under physiologic and disease conditions was supported by direct visualization and quantitative analysis.