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

2. Problem Statement
To measure capillary recruitment in vivo or in situ in the skeletal muscle microvascula-ture of mice in order to study muscle glucose uptake.

3. Muscle Glucose Uptake
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

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

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

6. Jablonski Diagram

7. Project Objective
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 insulin-stimulated MGU.

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

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

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

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