Cardiac tissue slices: Preparation, handling, and successful optical mapping

Wang, Ken; Lee, Peter; Mirams, Gary R.; Sarathchandra, Padmini; Borg, Thomas K.; Gavaghan, David J.; Kohl, Peter; Bollensdorff, Christian

Abstract

© 2015 the American Physiological Society. Cardiac tissue slices are becoming increasingly popular as a model system for cardiac electrophysiology and pharmacology research and development. Here, we describe in detail the preparation, handling, and optical mapping of transmembrane potential and intracellular free calcium concentration transients (CaT) in ventricular tissue slices from guinea pigs and rabbits. Slices cut in the epicardium-tangential plane contained wellaligned in-slice myocardial cell strands (“fibers”) in subepicardial and midmyocardial sections. Cut with a high-precision slow-advancing microtome at a thickness of 350 to 400 µm, tissue slices preserved essential action potential (AP) properties of the precutting Langendorff- perfused heart. We identified the need for a postcutting recovery period of 36 min (guinea pig) and 63 min (rabbit) to reach 97.5% of final steady-state values for AP duration (APD) (identified by exponential fitting). There was no significant difference between the postcutting recovery dynamics in slices obtained using 2,3-butanedione 2-monoxime or blebistatin as electromechanical uncouplers during the cutting process. A rapid increase in APD, seen after cutting, was caused by exposure to ice-cold solution during the slicing procedure, not by tissue injury, differences in uncouplers, or pH-buffers (bicarbonate; HEPES). To characterize intrinsic patterns of CaT, AP, and conduction, a combination of multipoint and field stimulation should be used to avoid misinterpretation based on source-sink effects. In summary, we describe in detail the preparation, mapping, and data analysis approaches for reproducible cardiac tissue slice-based investigations into AP and CaT dynamics.

Citation

Wang, K., Lee, P., Mirams, G. R., Sarathchandra, P., Borg, T. K., Gavaghan, D. J., …Bollensdorff, C. (2015). Cardiac tissue slices: Preparation, handling, and successful optical mapping. AJP - Heart and Circulatory Physiology, 308(9), H1112-H1125. https://doi.org/10.1152/ajpheart.00556.2014

• Journal Article Type: Article
• Acceptance Date: Jan 14, 2015
• Publication Date: May 1, 2015
• Deposit Date: Jan 14, 2020
• Publicly Available Date: Feb 28, 2020
• Journal: American Journal of Physiology-Heart and Circulatory Physiology
• Print ISSN: 0363-6135
• Electronic ISSN: 1522-1539
• Publisher: American Physiological Society
• Peer Reviewed: Peer Reviewed
• Volume: 308
• Issue: 9
• Pages: H1112-H1125
• DOI: https://doi.org/10.1152/ajpheart.00556.2014
• Public URL: https://nottingham-repository.worktribe.com/output/3217477
• Publisher URL: https://journals.physiology.org/doi/full/10.1152/ajpheart.00556.2014