Ice crystal growth in skeletal muscle fibres
- 1 September 1975
- journal article
- Published by Wiley in Journal of Microscopy
- Vol. 105 (1), 67-80
- https://doi.org/10.1111/j.1365-2818.1975.tb04037.x
Abstract
Ice crystal growth was studied in rapidly frozen skeletal muscle fibres which were treated with cryo‐protective additives (glycerol, DMSO, sucrose) or which were untreated. Freeze cleaving and etching was the basic method, with conventional plastic embedding and cryo‐ultramicrotomy as complementary techniques. Extensive crystal growth occurred during freezing in all unprotected fibres. Just below the fibre surface the crystals were numerous but small, while deeper in the fibre they were fewer but larger. The deeper within the specimen a fibre was located, the larger, in general, was the crystal size. The crystal volume density was about 55%, irrespective of crystal size. Ice recrystallization was practically absent at the temperature normally used in cryo‐sectioning (–70°C). Anti‐freeze treatment eliminated crystal growth. If the anti‐freeze agents were used in non‐toxic concentrations, however, their effect on crystal growth was very limited. ‘Dry’‐cut, freeze‐dried ultra‐thin cryosections of protected and unprotected fibres confirmed these observations, while sections obtained by ‘wet’ cryo‐cutting showed no apparent signs of crystal growth. In plastic sections of frozen and thawed fibres a previous occurrence of crystals was only slightly indicated. In interpreting the ultrastructure in ‘wet’‐cut cryo‐sections of unprotected frozen mucle fibres, the distorting effects of ice crystals through mechanical compression and alterations in sectioning conditions, must be taken into consideration. Crystal growth also strongly limits the possibilities of using ‘dry’‐cut sections of untreated frozen tissue for analytical electron microscopy; only the most superficial parts of the fibres seem to be suitable for microanalysis.Keywords
This publication has 40 references indexed in Scilit:
- Preparing sections of skeletal muscle for transmission electron analytical microscopy (TEAM) of diffusible elementsJournal of Microscopy, 1975
- Recent developments in analytical electron microscopyJournal of Microscopy, 1973
- A TECHNIQUE FOR ULTRACRYOTOMY OF CELL SUSPENSIONS AND TISSUESThe Journal of cell biology, 1973
- Myofilaments and cross bridges as demonstrated by freeze-fracturing and etchingJournal of Ultrastructure Research, 1972
- Relaxation of glycerinated muscle: Low-angle X-ray diffraction studiesJournal of Molecular Biology, 1972
- Structure of the myosin-containing filament assembly (A-segment) separated from frog skeletal muscleJournal of Molecular Biology, 1971
- Recent progress in the freeze-etching techniquePhilosophical Transactions of the Royal Society of London. B, Biological Sciences, 1971
- The low-angle X-ray diagram of vertebrate striated muscle and its behaviour during contraction and rigorJournal of Molecular Biology, 1967
- BIOLOGICAL IMPLICATIONS OF DMSO BASED ON A REVIEW OF ITS CHEMICAL PROPERTIES*Annals of the New York Academy of Sciences, 1967
- Low-Temperature Forms of Ice as Studied by X-Ray DiffractionNature, 1960