Generation of Porcine Ventricular Slices: A Vibratome-based Technique to Prepare Ultra-thin Slices of Pig Heart Tissue

Abstract

Source: Ou, Q. et. al., Slicing and Culturing Pig Hearts under Physiological Conditions. J. Vis. Exp. (2020)

This video demonstrates a vibratome-based procedure to generate viable, ultra-thin sections of pig heart. The prepared tissue slices can be cultured and used as model systems for pharmacological testing.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. Pig Heart Tissue Slicing

1. Set up the tissue bath on the vibratome, add ice to the tissue bath cooling jacket, then add Tyrode's solution into the tissue bath. Set up 1 L plastic jar to collect the disposal of melted ice from the tissue bath cooling jacket. For 1 L of the slicing Tyrode's solution, mix 3 g/L 2,3-butanedione monoxime (BDM), 140 mM NaCl (8.18 g), 6 mM KCl (0.447 g), 10 mM D-glucose (1.86 g), 10 mM HEPES (2.38 g), 1 mM MgCl₂ (1 mL of 1 M solution), 1.8 mM ....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Agarose	Bioline USA	BIO-41025
2,3-Butanedione monoxime (BDM)	Fisher	AC150375000
Calcium Chloride (CaCl2)	Fisher	C79-500
Ceramic Blades for Vibrating Microtome	Campden Instruments	7550-1-C
D-Glucose	Fisher	D16-1
Disposable Scalpel #20	Biologyproducts.com	DS20X
Falcon Cell Strainers, Sterile, Corning	VWR	21008-952
Heparin sodium salt	Sigma-Aldrich	H3149-50KU
HEPES	Fisher	BP310-1
Histoacryl BLUE Tissue glue	Amazon		https://www.amazon.com/HISTOACRYL-FLEXIBLE-1051260P-Aesculap-Adhesive/dp/B074WB5185/
Iris Spring scissors	Fisher	NC9019530
Iris Straight Scissors	Fisher	731210
Magnesium Chloride (MgCl2)	Fisher	M33-500
Oxygen regulator	Praxair
Oxygen tanks	Praxair
Plastic Pasteur pipettes	Fisher	13-711-48
Potassium Chloride (KCl)	Fisher	AC193780010
Razor rectangle blades	Fisher	12-640
Retractable scalpels	Fisher	22-079-716
Sodium Bicarbonate (NaHCO3)	Fisher	AC217125000
Sodium Chloride (NaCl)	Fisher	AC327300010
Vibrating Microtome	Campden Instruments	7000 SMZ-2