Abstract
Ultrathin frozen sections of biological tissues for electron microscopy provide certain advantages in cytochemical studies in which the penetration of cells by large molecules is necessary and in morphological studies of cellular constituents which are dissolved by the reagents employed in routine plastic embedding. The recent introduction of several types of commercially available cryo-ultramicrotomes makes it possible for many laboratories to employ this valuable tool. This paper summarizes recent improvements in the methods developed in this laboratory for preparing ultrathin frozen sections and reviews some of the inherent problems involved in their use. These procedures may serve as a baseline for other investigators who can then modify or adapt them for their specific purposes.
Full Text
The Full Text of this article is available as a PDF (1.9 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- BERNHARD W. ULTRAMICROTOMIE 'A BASSE TEMP'ERATURE. Annee Biol. 1965 Jan-Feb;41:5–19. [PubMed] [Google Scholar]
- Bernhard W., Leduc E. H. Ultrathin frozen sections. I. Methods and ultrastructural preservation. J Cell Biol. 1967 Sep;34(3):757–771. doi: 10.1083/jcb.34.3.757. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dollhopf F. L., Sitte H. Die Shandon-Reichert-Kühleinrichtung FC-150 zum Herstellen von Ultradüm- und Feinschnitten bei extrem niederen Temperaturen. I. Gerätetechnik. Mikroskopie. 1969;25:17–32. [PubMed] [Google Scholar]
- GILEV V. P. The use of gelatin for embedding biological specimens in preparation of ultrathin sections for electron microscopy. J Ultrastruct Res. 1958 Aug;1(4):349–358. doi: 10.1016/s0022-5320(58)90006-6. [DOI] [PubMed] [Google Scholar]
- Hodson S., Marshall J. Ultracryotomy: a technique for cutting ultrathin sections of unfixed frozen biological tissues for electron microscopy. J Microsc. 1970 Apr;91(2):105–117. doi: 10.1111/j.1365-2818.1970.tb02212.x. [DOI] [PubMed] [Google Scholar]
- Leduc E. H., Bernhard W., Holt S. J., Tranzer J. P. Ultrathin frozen sections. II. Demonstration of enzymic activity. J Cell Biol. 1967 Sep;34(3):773–786. doi: 10.1083/jcb.34.3.773. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Leduc E. H., Bernhard W. Recent modifications of the glycol methacrylate embedding procedure. J Ultrastruct Res. 1967 Jul;19(1):196–199. doi: 10.1016/s0022-5320(67)80068-6. [DOI] [PubMed] [Google Scholar]
- Luft J. H. Fine structures of capillary and endocapillary layer as revealed by ruthenium red. Fed Proc. 1966 Nov-Dec;25(6):1773–1783. [PubMed] [Google Scholar]
- MARINOZZI V. CYTOCHIMIE ULTRASTRUCTURALE DU NUCL'EOLE--RNA ET PROT'EINES INTRANUCL'EOLAIRES. J Ultrastruct Res. 1964 Jun;10:433–456. doi: 10.1016/s0022-5320(64)80021-6. [DOI] [PubMed] [Google Scholar]
- Napolitano L., Lebaron F., Scaletti J. Preservation of myelin lamellar structure in the absence of lipid. A correlated chemical and morphological study. J Cell Biol. 1967 Sep;34(3):817–826. doi: 10.1083/jcb.34.3.817. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pease D. C. Eutectic ethylene glycol and pure propylene glycol as substituting media for the dehydration of frozen tissue. J Ultrastruct Res. 1967 Nov;21(1):75–97. doi: 10.1016/s0022-5320(67)80007-8. [DOI] [PubMed] [Google Scholar]
- Zotikov L., Bernhard W. Localisation au microscope électronique de l'activité de certaines nucléases dans des coupes à congélation ultrafines. J Ultrastruct Res. 1970 Mar;30(5):642–663. doi: 10.1016/s0022-5320(70)90057-2. [DOI] [PubMed] [Google Scholar]