Indiana University, School of Medicine, Department of Medicine

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Biosensor Development Core

Time series of FRET measurements and images collected from the liver of a mouse expressing the protein kinase A FRET biosensor AKAR4.1 (delivered by adenovirus) following IV injection of glucagon (Reproduced from Tao et al., AJP Cell. 2015

Time series of 800nm/900nm multiphoton excitation ratio images of cells expressing the reactive oxygen sensor RO-GFP2 following addition of peroxide to their medium

Volumetric representation of calcium responses of GCamp6s expressed in cultured Ins-2 cells before and after addition of caffeine (Time is vertical)

The objective of the Core is to develop, characterize and validate fluorescence protein biosensors for intravital multiphoton microscopy.

The cloning and engineering of genetically encoded fluorescent proteins has allowed the development of many unique tools for studies in cell biology, medicine, and physiology. When combined with intravital microscopy, these probes have the potential to report events from inside living cells in the intact organism. However, the utility of the fluorescent protein biosensors for intravital microscopy has been limited by the poorly characterized behaviors of fluorescent proteins under two-photon excitation. The goal of the Multiphoton Biosensor Development core is to develop and rigorously validate a set of biosensor probes optimized for intravital multiphoton microscopy studies of renal function. Modular probes developed by the core will be incorporated into a toolkit of probes and protocols that will be provided to renal investigators. The Indiana OBrien Center is founded upon the mission of developing and implementing methods of microscopy that provide unique and powerful insights into renal function and dysfunction. The Multiphoton Biosensor Development Core will play a critical role in this mission, providing renal investigators with powerful new biosensor protocols for intravital microscopy, supporting unique studies addressing fundamental issues of renal physiology, cell biology and pathophysiology.