Snap Freezing of Muscle Tissue for Sectioning: A Protocol to Obtain Ultrathin Sections of Rapidly Frozen Murine Muscle Tissue

Abstract

Source: Bonetto, A. et al. The Colon-26 Carcinoma Tumor-bearing Mouse as a Model for the Study of Cancer Cachexia. J. Vis. Exp. (2016)

This video describes the procedure for snap freezing muscle tissue and obtaining its thin sections. The technique can help study cancer-related cachexia in colon cancer mouse models.

Protocol

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

1. Muscle Freezing and Mounting

1. Immerse the bottom half of a small plastic beaker (50 mL) containing isopentane into liquid nitrogen. The isopentane is ready to use when it becomes slightly viscous and forms a solid white laminate lining the inside of the beaker (temperature: -160 °C).
   NOTE: Always use non-sparking bronze or aluminum hand tools. Avoid breathing product vapor. Use with adequate ventilation. If dealing with a spill and ventila....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
OCT	Tissue-Tek	4583
Cryostat	Leica	CM1850
Cork disks	Electron Microscopy Sciences	63305
Superfrost plus glass slides	VWR	48311-703