Summary
An X-ray microanalytical and morphological investigation was carried out on rapidly frozen freeze-dried or freeze-substituted tissues. A comparison was made between different embedding and polymerisation procedures following freeze substitution and freeze drying. The investigation also included an analysis of specimens which had been infiltrated, embedded and polymerised by ultraviolet irradiation at low temperatures with Lowicryl-HM20. The method of freeze drying, followed by embedding and polymerisation at low temperatures in vacuo was found to give satisfactory results, comparable with more tedious and hazardous freeze substitution technique.
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References
Altmann R (1890) Die Elementarorganismen und ihre Beziehungen zu den Zellen. Veit, Leipzig
Anniko M, Lim D, Wróblewski R (1984) Elemental composition of individual cells and tissues in the cochlea. Acta Otolaryngol (in press)
Bensley RR, Gersh I (1933) Studies on cell structure by the freezedrying method. I. Introduction., II. The nature of the mitochondria in the hepatic cell of Amblystoma. Anat Rec 57:205–237
Carlemalm E, Graviato RM, Villinger W (1982) Resin development for electron microscopy and an analysis of embedding at low temperature. J Microsc 126:123–143
Edelmann L (1978) A simple freeze-drying technique for preparing biological tissue without chemical fixation for electron microscopy. J Microsc 112:243–248
Edelmann L (1980) Potassium binding sites in muscle. Electron microscopic visualization of K, Rb, and Cs in freeze-dried preparations and autoradiography at liquid nitrogen temperature using 86 Rb and 134 Cs. Histochemistry 67:233–242
Elfvin LG (1963) The ultrastructure of the plasma membrane and myelin sheat of peropherial nerve fibers after fixation by freezedrying. J Ultrastruct Res 8:283–304
Gersh I (1932) The Altmann technique for fixation by drying while freezing. Anat Rec 53:309–337
Hanzon V, Hermodson LH (1960) Freeze-drying of tissues for light and electron microscopy. J Ultrastruc Res 4:332–348
Hoerr NL (1936) cytological studies by the Altmann-Gersh freezedrying method. I. Recent advances in the technique. Anat Rec 65:293–317
Ingram FD, Ingram MJ (1980) Freeze-dried, plastic-embedded tissue preparation: a review. Scanning Electron Microsc IV:147–160
Ingram FD, Ingram MJ, Hogben CAM (1974) An analysis of the freeze-dried, plastic embedded electron probe specimen preparation. In: Hall T, Echlin P, Kaufman P (eds) Microprobe analysis as applied to cells and tissues. Academic Press, New York, pp 119–146
Marshall AT (1980) Freeze-substitution as a preparation technique for biological X-ray microanalysis. Scanning Electron Microsc II:395–408
Marshall AT (1980) Sections of freeze-substituted specimens. In: Hayat MA (ed) X-ray microanalysis in biology. University Park Press, Baltimore, pp 207–240
Monroe RG, Gamble WJ, La Farge CG, Gamboa R, Morgan CL, Rosenthal A, Bullivant S (1968) Myocardial ultrastructure in systole and diastole using ballistic cryofixation. J Ultruct Res 22:22–36
Richards HG, Steinback HB, Anderson TF (1943) Electron microscope studies of squid giant axoplasm. J Cell Comp Physiol 21:129–141
Scott GH (1933) A critical study and review of the method of microincineration. Protoplasma 20:133–151
Scott GH, Packer DM (1939a) The electron microscopy as an analytical tool for the localization of minerals in biological tissues. Anat Rec 74:17–30
Scott GH, Packer DM (1939b) An electron microscope study of magnesium and calcium in muscle. Anat Rec 74:31–46
Simpson WL (1941a) An experimental analysis of the Altmann technic of freeze-drying. Anat Rec 80:173–189
Simpson WL (1941b) The application of the Altmann method to the study of the Golgi apparatus. Anat Rec 80:329–345
Sjöstrand F (1943a) Fixering och preparing for elektronmikroskopisk undersökning av vävnad. Nord Med 19:1207–1222
Sjöstrand F (1943b) Electron-microscopic examination of tissues. Nature 151:725–726
Wróblewski R (1982) Healthy and diseased striated muscle studied by analytical scanning electron microscopy with special reference to fibre type. Scanning Electron Microsc III:1173–1189
Wróblewski R, Edström L (1983) Changes in elemental composition of single muscle fibers following tenotomy of the rat soleus muscle. Muscle Nerve 6:490–496
Wróblewski R, Edström L (1984) Recent applications of X-ray microanalysis in muscle pathology. Scanning Electron Microsc I:249–259
Wróblewski R, Jansson E (1975) Fine structure of single fibers of human skeletal muscle. Cell Tissue Res 161:471–476
Wróblewski J, Wróblewski R (1982) New preparation method for X-ray microanalysis of biological semithin sections. J Ultrastruct Res 81:375–376
Wróblewski J, Müller RM, Wróblewski R, Roomans GM (1983) Quantitative X-ray microanalysis of semi-thick cryosections. Histochemistry 77:447–463
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Wróblewski, R., Wróblewski, J. Freeze drying and freeze substitution combined with low temperature-embedding. Histochemistry 81, 469–475 (1984). https://doi.org/10.1007/BF00489752
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DOI: https://doi.org/10.1007/BF00489752