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Single Fiber Function Core

Single Fiber Function Core. Co-Directors: Ken Campbell and Karyn Esser Seed Funds from Center for Muscle Biology and ~5 Principal Investigators. Outline. Types of muscle we can use and how we prepare them Overview of the experimental approach

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Single Fiber Function Core

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  1. Single Fiber Function Core Co-Directors: Ken Campbell and Karyn Esser Seed Funds from Center for Muscle Biology and ~5 Principal Investigators

  2. Outline • Types of muscle we can use and how we prepare them • Overview of the experimental approach • What measurements can we make and what do they tell us? • How do I get started with the Core?

  3. What muscles can we use? • Almost anything - skeletal and cardiac • We have more experience with mammalian tissue but we can in principle do other things (e.g. lobster!) • Human samples • Prior KY experience with: rat psoas, rat soleus, rat diaphgram various murine skeletal muscles, murine diaphragm rat, murine, porcine and human myocardium

  4. Chemically Permeabilized Muscle Normal (Intact) muscle Permeabilized ('skinned') muscle

  5. Preparing Skeletal Muscle "Bundles"

  6. Preparing Skeletal Muscle "Bundles"

  7. Practicalities • We normally store skeletal 'bundles' at -20C for ~3 weeks Problems if you store longer than that • Potential to get longer-term storage with cryopreservation? • Cardiac tissue is simply ground up with a homogenizer. We often work with frozen tissue that has been in the freezer for months

  8. Outline • Types of muscle we can use and how we prepare them • Overview of the experimental approach • What measurements can we make and what do they tell us? • How do I get started with the Core?

  9. Chemically Permeabilized Muscle Normal (Intact) muscle Permeabilized ('skinned') muscle

  10. Muscle Experimental Setup

  11. Outline • Types of muscle we can use and how we prepare them • Overview of the experimental approach • What measurements can we make and what do they tell us? • How do I get started with the Core?

  12. Experimental Protocol

  13. Experimental Parameters and (simple) Interpretation

  14. Force / Velocity & Power Output

  15. 45 Year Old Female, LV ExplantFamilial Dilated Cardiomyopathy Campbell, Mitov, Ferreira, Shekar and Bonnell (Unpublished)

  16. Short-range force increases with stretch velocityin human myocardium pCa 5.9 ~10% Maximal Activation Campbell, Mitov, Ferreira, Shekar and Bonnell (Unpublished)

  17. Outline • Types of muscle we can use and how we prepare them • Overview of the experimental approach • What measurements can we make and what do they tell us? • How do I get started with the Core? Speak to Ken k.s.campbell@uky.edu

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