Dr. Aron Geurts Profile

Dr. Aron Geurts

at Medical College of Wisconsin

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Publications (2)

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Proceedings Article | 5 March 2021 Presentation + Paper

Assessing drug-cell and drug-tissue interactions through spectral FRET imaging

Silas Leavesley, Naga Annamdevula, Aron Geurts, D. J. Pleshinger, Thomas Rich

Proceedings Volume 11624, 116240D (2021) https://doi.org/10.1117/12.2583249

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Structural complexity and heterogeneity may play critical roles in pathophysiology and therapeutic effect, at length scales ranging from subcellular (nm) to whole organ (cm). Here, we report on efforts to utilize a multidimensional spectral FRET imaging approach for visualization of the effects of pharmacologic treatments on tissues. We have developed a transgenic rat line expressing a cAMP FRET reporter for visualization of second messenger signaling in tissues in response to treatment and pathological conditions. When utilized with vascular and lung preparations, transgenic FRET tissues displayed the ability to elicit agonist-induced cAMP responses in single cells.

Proceedings Article | 5 March 2021 Presentation + Paper

Hyperspectral imaging approaches to characterize the distribution and expression of FRET in tissues isolated from a novel transgenic rat expressing a FRET-based probe

Naga Annamdevula, Aron Geurts, Silas Leavesley, Thomas Rich

Proceedings Volume 11649, 1164904 (2021) https://doi.org/10.1117/12.2583267

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Fӧrster resonance energy transfer (FRET) based cAMP sensors have become a standard approach to visualize cAMP signals in cells. However, FRET sensors inherently offer low signal-to-noise ratios, making cAMP measurements challenging, especially in 3 spatial dimensions. In previous studies, we used 4-dimensional (x, y, z, and λ) hyperspectral imaging and analysis approaches to measure agonist – induced distribution of cAMP signals in cultured pulmonary microvascular endothelial cells. In many cell types including PMVECs, transient transfection of FRET sensors results in a low transfection efficiency (1-5%), which limits the possibilities of studying spatial distribution of FRET signals in many cells simultaneously. We developed a transgenic rat model that expresses the H187 FRET sensor. In current studies, we utilize hyperspectral imaging and analysis approaches to characterize the distribution of FRET signals in different tissues and vessels harvested from transgenic rats. Hyperspectral z-stacks of tissues and vessels were acquired using a Nikon A1R confocal microscope equipped with 20X multi-immersion objective and a 32 channel PMT detector. Samples were excited using 405 nm and 561 nm lasers and emission was collected from 424 nm -724 nm at 10 nm intervals. Tissues including lungs, heart, mesentery, kidney, expressed the H187 FRET sensor. In conclusion, transgenic rats provide a platform to visualize cAMP signals in-vitro and in-situ.

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