Sample, Imaging, Metabolomics and Proteomics (SIMP) Core

University Of Colorado Denver, Aurora CO

Investigators

Linked publications, trials & patents

Abstract

SIMP CORE ABSTRACT: The capabilities of this technical Core predicate the kinds of questions we can address in our projects. Our strategies emphasize biophysical methods, to process large numbers of human samples run consistently and quantitatively. These approaches expand on our previous successes with imaging molecular colocalization in tissues and proteomics. In this cycle we extend antibody based detection approaches such as imaging/cytometry/ELISA, to mass spectrometry (MS) based metabolomics, affinity purification mass spectrometry (AP-MS) for protein:protein discovery and stable isotope labeling for flux analysis and absolute quantification. This Core centralizes various animal experimentation and animal care protocols to provide uniform and reproducible quality for all projects. These animal models provide the critical bridge between in vitro studies and clinical findings that is necessary to complete the missions of our translational research. We have adopted bar coding and sample aliquot cataloguing, as practiced by HS core, to store tissues and biofluids from animal experiments. Over the past cycle, we have installed and implemented state-of-art MS metabolomics capabilities and established productivity. Metabolites offer a valuable readout for phenotype, as they are the end product of catalytic events that are modulated at multiple levels (transcription, modification, protein turn-over, etc.). Stable isotope flux analysis and the use of a wide array of available metabolic enzymes inhibitors allow for detailed mechanistic studies. Above and beyond, processing microliter volumes of hundreds to thousands of samples in a timely fashion, this Core must address emerging bottlenecks and develop solutions in advance. The Core Leaders Drs. Banerjee and Hansen have identified several such areas including fast point-of care-metabolomics, label free imaging (e.g. with 2nd harmonics) and improved imaging of fibrinolytic processes in whole blood under physiological flow that are being explored. Impact: The goals of this Core are to provide projects with the animal models and analytical procedures necessary to test hypotheses and discover new mechanisms of adaptive and maladaptive responses to trauma and hemorrhagic shock. All of this is aimed at the ultimate goal of improving personalized diagnosis and care for the individual patient. 1