The Integrative and Applied Muscle Biology (IAMB) Laboratory, located in MDC 3026, is fully equipped to study muscle contractility at a variety of levels conduct all of the biochemical assays related to the proposed research. In addition to typical wet lab apparatus (e.g., dedicated pH meter, -40 and -80 C freezers, refrigerated centrifuge), the IAMB Laboratory is equipped to carry out tissue biochemistry (e.g., immunoblotting, SDS-PAGE). Specific equipment related to muscle testing includes:
In Situ Contractile Testing Apparatus: The PI has a customized Kopf Spinal Frame which can be fitted with various World Precision Instruments force transducers, a personal computer for data storage and analysis, 1 Grass S48 electrical stimulator with PSIU6 stimulus isolation unit, a WPI TB4M multichannel Amplifier, a heat lamp and aBAT-12 temperature monitoring system with rectal probe. This system has been used in several peer-reviewed publications (PMID#24316040, #21993787, #21966643, #20874807, #16857718, #18091018) for in situ contractile testing of the rat hindlimb. A VetEquip Compact Anesthesia System (isoflurane) is available from LAR.
Isolated Muscle Testing System: The PI has an Aurora 1205A Isolated Muscle System interfaced with Spike 2 software, CED Micro1401 interface, circulating water system for temperature control, Grass S48 electrical stimulator & CEM stimulus splitter for the collection of ex vivo contractile data. The interchangeable components of the system have been customized to allow testing of both rat and mouse muscles. Appropriate CO2 and O2 cylinders are available for the organ baths.
JASCO FP6200 Fluorometer: This cuvette based system allows collection of spectral and time series data for metabolite and enzymatic activity assays. The dual excitation and dual emission detection of fluorescent dyes such as those used to measure calcium handling, as well as . For the proposed studies, we will be working with Fura-2 in our assays of SR calcium leak, release and re-uptake. The system also has a magnetic stirring module and water-jacketed cuvette temperature controller. The system is also capable of performing fluorometric metabolite and enzyme assays.
Grip Testing Apparatus: The PI has a Columbus Instruments dual sensor 1027DR grip strength meter with a triangle bar, T-bar and grid attachments.