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Measurement of Capillary Recruitment

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

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Measurement of Capillary Recruitment

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

  2. 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.

  3. Regulation of Microcirculation QM= Blood Flow to Muscle QC= Blood Flow per Capillary NC= Number of Perfused Capillaries VC= Blood Velocity AC= Capillary Cross Sectional Area QM=QCNC=VCACNC • Blood flow to the muscles is directly proportional to the number of perfused capillaries (NC) • Surface area for glucose exchange is proportional to NC, so a higher blood velocity will not be as efficient

  4. Capillary Recruitment • An increase in the number of perfused capillaries is known as capillary recruitment • 10% of capillaries are perfused in resting skeletal muscles • Capillary density is a function of metabolic requirements

  5. 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

  6. 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.

  7. 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

  8. Jablonski Diagram

  9. Methods • 1.1 ug/mL saline solution of TX Red-Dextran T70 infused at 1.375 uL/min • Dye solution equilibrated in mouse after 100 min • Catheterized mouse anesthetized using Nambitol • Surgery performed, vasus lateralus muscle exposed on right leg • Mouse mounted on confocal microscope • 50 uL sodium nitroproside (SNP) injected to vasodilate • Observations made and images collected

  10. Results • “Honeycomb” structure imaged • Capillary flow evident in structure

  11. Capillary Images

  12. 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

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

  14. Future Plans • Test Fluorescent Microscopy on 2/16/01 using the confocal microscope and implemented technique. • Adapt this technique to study MGU. • If adaptation unsuccessful, use Optical Coherence Tomography (OCT)

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

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