Preservation of shock-frozen myocardial tissue as shown by cryo-ultramicrotomy and freeze-fracture studies. 1977

T S Saetersdal, and J Røli, and R Myklebust, and H Engedal

Myocardial tissue was quickly quench-frozen by letting the specimen fall either into liquid nitrogen (LN2) or into LN2-slush, or, by drill-propelling the specimen into LN2 at a speed of 1.5 m/s. The preservation of the tissues was studied in ultrathin, dry-cut and freeze-dried cryo-sections. Shock-freezing by propelling into LN2 yields extensive areas of well preserved tissue without hole damage. Quenching into LN2-slush shows variable results, while a maximum of hole damage is obtained by letting the specimen fall into LN2. Replicas of freeze-fractured tissue which had been quench-frozen by the same methods confirmed these observations. By the drill-propelling, areas 2000-5000 micrometer2 wide reveal good preservation without visible ice crystals. Such areas occur within a superficial band of tissue to a depth of c. 20 micrometer. Even at 35 micrometer and 50 micrometer depth good preservation may be registered at the periphery of the replicas. Description is given of modifications in accessories and methods in order to make the LKB CryoKit more suitable as a routine instrument. These modifications cover replacement of the knife coolant container by a coolant brass container in open connection with three copper tubes, replacement of one of the LKB Dewar flasks by a 25 litre Dewar equipped with a Balzers filling device, and replacement of the LKB plastic sleeve fixed to the back of the specimen coolant container, by two perspex-glass discs mounted in the slit between the Ultrotome base and the cryo-chamber. In addition, modified constructions are presented of the grid-carrier, the press-assembly as well as of the container for freeze-drying and warming up of the frozen sections.

UI MeSH Term Description Entries
D008867 Microtomy The technique of using a microtome to cut thin or ultrathin sections of tissues embedded in a supporting substance. The microtome is an instrument that hold a steel, glass or diamond knife in clamps at an angle to the blocks of prepared tissues, which it cuts in sections of equal thickness. Thin Sectioning,Ultramicrotomy,Sectioning, Thin,Sectionings, Thin,Thin Sectionings
D009206 Myocardium The muscle tissue of the HEART. It is composed of striated, involuntary muscle cells (MYOCYTES, CARDIAC) connected to form the contractile pump to generate blood flow. Muscle, Cardiac,Muscle, Heart,Cardiac Muscle,Myocardia,Cardiac Muscles,Heart Muscle,Heart Muscles,Muscles, Cardiac,Muscles, Heart
D005614 Freeze Fracturing Preparation for electron microscopy of minute replicas of exposed surfaces of the cell which have been ruptured in the frozen state. The specimen is frozen, then cleaved under high vacuum at the same temperature. The exposed surface is shadowed with carbon and platinum and coated with carbon to obtain a carbon replica. Fracturing, Freeze,Fracturings, Freeze,Freeze Fracturings
D005629 Frozen Sections Thinly cut sections of frozen tissue specimens prepared with a cryostat or freezing microtome. Frozen Section,Section, Frozen,Sections, Frozen
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D051379 Mice The common name for the genus Mus. Mice, House,Mus,Mus musculus,Mice, Laboratory,Mouse,Mouse, House,Mouse, Laboratory,Mouse, Swiss,Mus domesticus,Mus musculus domesticus,Swiss Mice,House Mice,House Mouse,Laboratory Mice,Laboratory Mouse,Mice, Swiss,Swiss Mouse,domesticus, Mus musculus

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