Measurement of Capillary Recruitment Stephanie Beatrous Snehal Patel

Measurement of Capillary Recruitment Stephanie Beatrous Snehal Patel Samar Sharma Advised by: David Wasserman, Ph. D.

Problem Statement n To measure capillary recruitment in vivo or in situ in the skeletal muscle microvasculature of mice in order to study muscle glucose uptake.

Muscle Glucose Uptake n Muscle Glucose Uptake (MGU) is a linear sequence of three processes. – – – Glucose delivery from blood to the interstitial space Glucose transport from interstitial space to the intercellular environment. Glucose phosphorylation by Hexokinase II

Significance of MGU Glucose plays a role in cell function by serving as an energy source. n Each of the three processes in MGU determine whether cellular needs for energy source are met. n As MGU is controlled by many physiological subsystems, alterations could lead to pathology. n

Diabetes n Diabetes is fundamentally a disorder of MGU. – – 16 million Americans 98 billion dollars in costs 18% of deaths (>25 yrs old). 7 th leading cause of death in 1993 – pharminfo. com

Jablonski Diagram

Project Objective n To design a technique to measure capillary recruitment in vivo or in situ in the skeletal muscle microvasculature of mice for the purpose of investigating basal and insulinstimulated MGU.

Past Work n Research of current imaging techniques – – – – – Optical Coherence Tomography (OCT) Intravital Microscopy Microspheres Functional Magnetic Resonance Imaging (f. MRI) Fluorescent Microscopy Two-photon Excitation Microscopy Whole-Field Microscopy Widefield Imaging Confocal Microscopy

Current Status n Research almost complete – n Need to continue assessing feasibility of methods Most Promising method identified – Fluorescent Microscopy

Future Plans Test Fluorescent Microscopy on 1/30/01 in Dr. Haselton’s lab at VU BME dept. n Adapt this technique to study MGU. n – – – – Signal to Noise ratio Spatial resolution Temporal resolution Fluorescent label(s) Light source wavelength(s) anesthetic vasodilator

Future Plans (cont. ) Test variations in the technique n Refine protocol for measuring capillary recruitment n Obtain images and assess quality of results n