Nuclear-Cytoplasmic Relationships in Human Cells in Tissue Culture

Micou, Julie; Collins, Carter C.; Crocker, T. Timothy

doi:10.1083/jcb.12.1.195

Cited by 14 publications

(3 citation statements)

References 7 publications

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“…Rapid cooling of the embedded specimen after full polymerization facilitated the separation of the cell support from the polymerized resin to a certain extent (Howatson & Almeida, 1958;Brinkley et al, 1967;Flaxman et al, 1969). T o this purpose also various coatings of glass cell supports have been employed, such as carbon (Bloom, 1960;Robbins & Gonatas, 1964), collagen (Heyner, 1963), silicone (Micou et al, 1962;Rosen, 1962), teflon (Chang, 1971) and polytetrafluorethane (Buckley, 1971).…”

Section: R E S U L T S a N D Discussionmentioning

confidence: 99%

“…In the literature, several marking techniques have been described to designate individual living cells for electronmicroscopic studies (Bloom, 1960;Micou et al, 1962;Robbins & Gonatas, 1964;Price, 1967;Buckley, 1971). These techniques all employed diamond markers in combination with glass cell supports and enabled studies of single cells in a plane planparallel to the cell support.…”

Section: R E S U L T S a N D Discussionmentioning

confidence: 99%

“…Techniques which permit comparative light and electronmicroscopic studies of cell cultures and cell spreads are of considerable value for experimental cell biology. Although a variety of methods in this field has been reported, only a few methods concern the electronmicroscopic aspects of single cells, individually selected in cell cultures in the living state (Bloom, 1960;Micou et al, 1962;Robbins & Gonatas, 1964, Price, 1967Buckley, 1971). Several technical problems exist in the electronmicroscopic studies of in vitro cultured cells, especially related to the separation of cells and cell support after polymerization of the embedding resin and the relocation of one cell of particular interest after embedding and polymerization.…”

Section: Introductionmentioning

confidence: 99%

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A new method for comparative light and electron microscopic studies of individual cells, selected in the living state

Ewijk

Hösli

1975

Journal of Microscopy

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SUMMARY A method is described which permits comparative light and electronmicroscopic studies of cell cultures, cell spreads or single selected cells which have been kept in the Plastic Film Dish (PFD). The PFD is a versatile large surface tissue culture chamber which, for electron microscopy, is mounted with a transparent FEP‐Teflon film bottom. Cells are observed, selected and marked on the PFD‐bottom with a high power inverted light microscope. The cells are fixed and dehydrated with a semi‐automatic device while they are still in situ in the PFD. During the preparation steps for electron microscopy the topographical relationship between individual cells and between cells and cell support is accurately retained. After embedding and polymerization the Teflon film is easily peeled off the polymerized Epon, leaving a replica of the mark around the selected cell. This permits relocation of the selected cells for ultrathin sectioning in a plane plan‐parallel to the original cell support. To enable orientated sectioning of selected cells in a plane perpendicular to the cell support, cells are tagged with Letraset‐letters after original embedding and polymerization. Subsequently the re‐embedded polymerized specimens are orientated in the microtome in a position which permits controlled thin sectioning of the tagged cells in the previously selected plane.

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