Abstract
Diethylene glycol distearate is used as a removable embedding medium to produce embeddment -free sections for transmission electron microscopy. The easily cut sections of this material float and form ribbons in a water-filled knife trough and exhibit interference colors that aid in the selection of sections of equal thickness. The images obtained with embeddment -free sections are compared with those from the more conventional epoxy-embedded sections, and illustrate that embedding medium can obscure important biological structures, especially protein filament networks. The embeddment -free section methodology is well suited for morphological studies of cytoskeletal preparations obtained by extraction of cells with nonionic detergent in cytoskeletal stabilizing medium. The embeddment -free section also serves to bridge the very different images afforded by embedded sections and unembedded whole mounts.
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- Capco D. G., Jeffery W. R. Differential distribution of poly(A)-containing RNA in the embryonic cells of Oncopeltus fasciatus. Analysis by in situ hybridization with a [3H]poly(U) probe. Dev Biol. 1978 Nov;67(1):137–151. doi: 10.1016/0012-1606(78)90305-6. [DOI] [PubMed] [Google Scholar]
- Capco D. G., Jeffrey W. R. Transient localizations of messenger RNA in Xenopus laevis oocytes. Dev Biol. 1982 Jan;89(1):1–12. doi: 10.1016/0012-1606(82)90288-3. [DOI] [PubMed] [Google Scholar]
- Capco D. G., Jäckle H. Localized protein synthesis during oogenesis of Xenopus laevis: analysis by in situ translation. Dev Biol. 1982 Nov;94(1):41–50. doi: 10.1016/0012-1606(82)90066-5. [DOI] [PubMed] [Google Scholar]
- Capco D. G., Penman S. Mitotic architecture of the cell: the filament networks of the nucleus and cytoplasm. J Cell Biol. 1983 Mar;96(3):896–906. doi: 10.1083/jcb.96.3.896. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Capco D. G., Wan K. M., Penman S. The nuclear matrix: three-dimensional architecture and protein composition. Cell. 1982 Jul;29(3):847–858. doi: 10.1016/0092-8674(82)90446-9. [DOI] [PubMed] [Google Scholar]
- Graham E. T. Improved diethylene glycol distearate embedding wax. Stain Technol. 1982 Jan;57(1):39–43. doi: 10.3109/10520298209066518. [DOI] [PubMed] [Google Scholar]
- Heuser J. E., Kirschner M. W. Filament organization revealed in platinum replicas of freeze-dried cytoskeletons. J Cell Biol. 1980 Jul;86(1):212–234. doi: 10.1083/jcb.86.1.212. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pardue R. L., Brady R. C., Perry G. W., Dedman J. R. Production of monoclonal antibodies against calmodulin by in vitro immunization of spleen cells. J Cell Biol. 1983 Apr;96(4):1149–1154. doi: 10.1083/jcb.96.4.1149. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Penman S., Fulton A., Capco D., Ben Ze'ev A., Wittelsberger S., Tse C. F. Cytoplasmic and nuclear architecture in cells and tissue: form, functions, and mode of assembly. Cold Spring Harb Symp Quant Biol. 1982;46(Pt 2):1013–1028. doi: 10.1101/sqb.1982.046.01.094. [DOI] [PubMed] [Google Scholar]
- REYNOLDS E. S. The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. J Cell Biol. 1963 Apr;17:208–212. doi: 10.1083/jcb.17.1.208. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rindler M. J., Chuman L. M., Shaffer L., Saier M. H., Jr Retention of differentiated properties in an established dog kidney epithelial cell line (MDCK). J Cell Biol. 1979 Jun;81(3):635–648. doi: 10.1083/jcb.81.3.635. [DOI] [PMC free article] [PubMed] [Google Scholar]
- SALAZAR H. DIETHYLENE GLYCOL DISTEARATE EMBEDDING AND ULTRAMICROTOME SECTIONING FOR LIGHT MICROSCOPY. Stain Technol. 1964 Jan;39:13–17. doi: 10.3109/10520296409061202. [DOI] [PubMed] [Google Scholar]
- SHALLA T. A., CARROLL T. W., DEZOETEN G. A. PENETRATION OF STAIN IN ULTRATHIN SECTIONS OF TOBACCO MOSAIC VIRUS. Stain Technol. 1964 Sep;39:257–265. doi: 10.3109/10520296409061241. [DOI] [PubMed] [Google Scholar]
- Singer R. H., Ward D. C. Actin gene expression visualized in chicken muscle tissue culture by using in situ hybridization with a biotinated nucleotide analog. Proc Natl Acad Sci U S A. 1982 Dec;79(23):7331–7335. doi: 10.1073/pnas.79.23.7331. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Taleporos P. Diethylene glycol distearate as an embedding medium for high resolution light microscopy. J Histochem Cytochem. 1974 Jan;22(1):29–34. doi: 10.1177/22.1.29. [DOI] [PubMed] [Google Scholar]
- Taleporos P. The composition and cutting properties of diethylene glycol distearate from various sources. J Histochem Cytochem. 1976 Dec;24(12):1285–1286. doi: 10.1177/24.12.1002977. [DOI] [PubMed] [Google Scholar]
- Wolosewick J. J., Porter K. R. Microtrabecular lattice of the cytoplasmic ground substance. Artifact or reality. J Cell Biol. 1979 Jul;82(1):114–139. doi: 10.1083/jcb.82.1.114. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wolosewick J. J., Porter K. R. Stereo high-voltage electron microscopy of whole cells of the human diploid line, WI-38. Am J Anat. 1976 Nov;147(3):303–323. doi: 10.1002/aja.1001470305. [DOI] [PubMed] [Google Scholar]
- Wolosewick J. J. The application of polyethylene glycol (PEG) to electron microscopy. J Cell Biol. 1980 Aug;86(2):675–661. doi: 10.1083/jcb.86.2.675. [DOI] [PMC free article] [PubMed] [Google Scholar]