Sample preparation

Ultramicrotome RMC PowerTome (Boeckeler Instruments), diamond knife (Diatome AG, Švajčiarsko), vibratome 7000smz (Campden Instruments), perfusion setup Back

Contact: RNDr. Marta Novotová, CSc., RNDr. Ivan Zahradník, CSc.

For electron-microscopic examination of the ultrastructure, tissue samples are embedded into the polymeric resin Durcupan and used for preparation of ultrathin sections (58 – 60 nm) in an ultramicrotome. Ultrathin sections are prepared using a diamond knife. Ultrathin sections, deposited on copper grids covered by a formvar membrane are used for examination of the samples in electron microscope.

Perfusion apparatus for chemical fixation of isolated working heart of experimental animals (rat, mouse) consists of three perfusion columns. Column #1 is filled with physiological saline and serves for removing blood from the working heart; column#2, containing calcium-free physiological saline is used for relaxation of the myocardium; and column #3 filled by fixation solution ensures prefixation of the whole myocardium.

Perfusion apparatus for isolation of cardiac myocytes from the left ventricle of the heart consists of three perfusion columns. Column #1 is filled with physiological saline and serves for removing blood from the working heart; column #2, containing calcium-free physiological saline is used for relaxation of the myocardium; and column #3 filled by enzyme solution serves for separation of individual cells.

The vibratome is used for preparation of slices from live tissue. Thin sections (10 – 450 µm) are prepared using a knife with stainless steel blade or with a ceramic blade from ultra hard zirconium.