Freeze fracture through the cytoskeleton, nucleus and nuclear matrix of lymphocytes studied by scanning electron microscopy

Haggis, G. H.; Schweitzer, Irwin; Hall, R. H.; Bladon, Trevor

doi:10.1111/j.1365-2818.1983.tb04271.x

Cited by 11 publications

(11 citation statements)

References 8 publications

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“…In agreement with work previously reported [1,5,6,12,16,25], the present work shows that the predominant intranuclear structures are represented by 30-nm diam chromatin fibers of indeterminate lengths. While these fibers and nuclear spaces [16], as well as some detail of the nuclear envelope, may be discerned in unexpanded cells ( Fig. 1 a, b), there emerges a clear and dramatic increase in resolution associated with hypotonic dispersal of cytoplasm and nucleoplasm (Figs..~ c, d and 2).…”

Section: Interphase Chromatinsupporting

confidence: 93%

“…Chromatin fibers positioned in the area subjacent to putative nuclear pores or within nuclear spaces [16], communicating with nuclear pores, were frequently found to present in an apparently more decondensed conformation than chromatin domains remote from the nuclear envelope, where this more open chromatin conformation was rarely observed. In fact, in regions underlying nuclear pores, nucleosomes comprising the chromatin fibers were frequently resolved as clusters (Figs.…”

Section: Interphase Chromatinmentioning

confidence: 99%

“…Specifically, chromatin fibers 30 nn'~ in diameter, ,~tie chromatosomes or nucleosomes comprisl~.g these fibers, the double membrane of the nuclear envelope including nuclear pores, as well as nuclear spaces [16], the potential spaces not discernible in unexpanded nuclei, were prominent after hypotonic expansion.…”

Section: Interphase Chromatinmentioning

confidence: 99%

“…While high-resolution scanning electron microscopy (SEM) has previously been used to examine chromatin [19], it has commonly been carried out on isolated nuclei or isolated chromatin [I, 21,36] or, alternatively, on detergent-extracted or deep-etched cells [I0- 16].…”

Section: Introductionmentioning

confidence: 99%

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Chromatin and nuclear envelope of freeze‐fractured, neuronal interphase nuclei, resolved by scanning electron microscopy

Boni

1988

Biology of the Cell

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High-resolution scanning electron microscopy (SEM) has previously been used to study intracellular detail, including chromatin. It has, however, been commonly carried out either on cellular subfractions or following extraction methods to visualize detail. In the work presented here, intracellular detail of neurons of the dorsal root was visualized in situ by viewing freeze-fracture faces obtained after hypotonic expansion. This procedure permits the detailed resolution, by SEM, of juxtanuclear and intranuclear detail to a degree impossible without hypotonic dispersal. In agreement with work previously reported, nuclear chromatin of these interphase cells presents largely as 30-nm fibers, with a next higher hierarchical structure imparted by swelling in magnesium chloride. Detailed analyses showed that particles as small as 10-nm nucleosomes comprising the 30-nm chromatin fiber could be resolved, with "end-on" views of such fibers showing 5 nucleosomes per helical turn of the fiber. Chromatin fibers positioned subjacent to nuclear pores, or associated with "nuclear spaces" communicating with nuclear pores, were frequently found to be resolved as clusters, in an apparently more decondensed conformation, rather than tightly coiled into the 30-nm fiber. In addition, details of the nuclear envelope, including nuclear pores and perinuclear filaments as well as membranes of the endoplasmic reticulum, decorated with ribosomes, were clearly resolved.

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Section: Interphase Chromatinsupporting

confidence: 93%

Section: Interphase Chromatinmentioning

confidence: 99%

Section: Interphase Chromatinmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Chromatin and nuclear envelope of freeze‐fractured, neuronal interphase nuclei, resolved by scanning electron microscopy

Boni

1988

Biology of the Cell

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“…The diameter of the cytoskeletal elements is not a reliable measure in scantling electron microscope studies, when the type of each element has to be identified, even when the thickness of the metal-coating is well under control (24). The present paper will therefore not discuss the relative contribution by the various cytoskeletal elements.…”

Section: Discussionmentioning

confidence: 95%

High resolution scanning electron microscopy of the subplasmalemmal cytoskeleton in human odontoblasts

Sögaard-Pedersen

Boye

1990

European J Oral Sciences

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– The subplasmalemmal cytoskeleton in human odontoblasts was studied by high resolution scanning electron microscopy. The odontoblast layer was isolated and exposed to formaldehyde, glutaraldehyde, and OsO4 for some specimens, while the membraneous structures and soluble proteins in the dental tissue were removed by Zenker's solution and 1% OsO4 for other specimens, without further fixation of the remaining components. The cytoskeletal elements comprised a dense network of interlacing filaments of different diameters in the cell body. Most cytoskeletal elements were parallel to the axis of the cell processes situated inside the dentinal tubules. The appearance and orientation of the investigated subplasmalemmal cytoskeletal elements was unaffected by the choice of method. Both methods confirm the presence of a subplasmalemmal cytoskeleton in human odontoblasts.

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Use of Peldri II (a fluorocarbon solid at room temperature) as an alternative to critical point drying for biological tissues

Kennedy

Williams

Gray³

1989

J. Elec. Microsc. Tech.

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A new chemical, Peldri II, is evaluated as a compound for drying soft biological tissues for scanning electron microscopy. Peldri II, a fluorocarbon, is a solid at room temperature and is a liquid above 25 degrees C. Cells or tissues are embedded in Peldri II by immersing them in the liquid form and allowing it to solidify. Once solidified, Peldri II will sublime with or without vacuum to dry tissues, probably without introducing surface tension. Several types of cells and tissues have been examined to compare preservation with Peldri II and critical point drying techniques. No differences were detected between the two techniques when normal surface structures were examined. Peldri II appears to be a significant improvement over hexamethyldisilazane as a drying agent for scanning electron microscopy. It is also very convenient for drying large numbers of samples.

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Freeze fracture through the cytoskeleton, nucleus and nuclear matrix of lymphocytes studied by scanning electron microscopy

Cited by 11 publications

References 8 publications

Chromatin and nuclear envelope of freeze‐fractured, neuronal interphase nuclei, resolved by scanning electron microscopy

Chromatin and nuclear envelope of freeze‐fractured, neuronal interphase nuclei, resolved by scanning electron microscopy

High resolution scanning electron microscopy of the subplasmalemmal cytoskeleton in human odontoblasts

Use of Peldri II (a fluorocarbon solid at room temperature) as an alternative to critical point drying for biological tissues

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