A new approach to the study of cell organelles with the electron microscope

Pihl, Erik; Bahr, G. F.

doi:10.1016/0014-4827(70)90644-0

Cited by 12 publications

(3 citation statements)

References 4 publications

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“…The nuclei and erythrocytes dehydrated by the routine ethanol method were indistinguishable, by SEM, from those dehydrated using the ethylene glycol-Cellosolve method of Pihl & Bahr (1970). Erythrocytes are osmotically sensitive cells and the quality of their ultrastructural preservation is highly dependent upon the method of fixation and dehydration (Brown, 1975).…”

Section: R E S U L T S and Discussionmentioning

confidence: 98%

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Cell and organelle shrinkage during preparation for scanning electron microscopy: effects of fixation, dehydration and critical point drying

Gusnard

Kirschner

1977

Journal of Microscopy

115

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The critical point drying method of preparing samples for scanning electron microscopy is associated with a variable amount of specimen shrinkage. We studied the causes of this phenomenon is isolated mouse hepatocyte nuclei and in human erythrocytes and found that the critical point drying process itself caused most of the shrinkage that we observed (a 25-30% reduction in diameter in both specimens). Glutaraldehyde fixation and ethanol dehydration caused only minimal size reduction, prior to critical point drying. Substitution of an inert (ethylene glycol-ethylene glycol monethyl ether) dehydration technique did not alter the final result. Previous studies in our laboratory using high resolution SEM and correlative transmission microscopy of isolated nuclei have demonstrated that the shrinkage represents a miniaturization of the organelles in which all structural components retain their usual relationships.

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Section: R E S U L T S and Discussionmentioning

confidence: 98%

“…! Bahr (1970). This method employs ethylene sow follorved by Cellosolve (ethylene glycol monoethyl ether; Fisher Scientific ocytes, adherent to formvar-coated glass SEM specimen discs, were processed erFding to the scheme in Fig.…”

Section: A T E R I a L S A N D M E T H O D Smentioning

confidence: 99%

Cell and organelle shrinkage during preparation for scanning electron microscopy: effects of fixation, dehydration and critical point drying

Gusnard

Kirschner

1977

Journal of Microscopy

115

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“…Dehydration of the cells was attempted by several methods, including treatment with a graded series of alcohol or acetone and treatment with acidified dimethoxypropane (13) . The best results were obtained by treating the cells with ethylene glycol at concentrations of 30, 60, 95, and 100% for3-5 min/concentration, followed by two 5-min rinses in 100% ethylene glycol monomethyl ether (14). The cells sank rapidly in 30 and 60% ethylene glycol, but centrifugation at 1,000 g for l min was required to recover the cells from 95 and 100% ethylene glycol .…”

Section: Methodsmentioning

confidence: 99%

Surface structure changes of rat adipocytes during lypolysis stimulated by various lypolysis stimulated by various lypolytic agents

Smith¹

1980

The Journal of Cell Biology

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A qualitative and quantitative electron microscopic study was performed on rat adipocytes during stimulation of lipolysis by various agents . Scanning electron microscopy of control cells revealed a spherical cell with a textured glycocalyx surface exhibiting small irregular projections. Globular surface evaginations or protrusions measuring 8-18 p,m in diameter were seen on cell hemispheres, and there was an average of one protrusion for every two hemispheres examined . Distribution analysis showed that 60% of the hemispheres had no protrusions, and 25, 10, and 5% of the hemispheres had one, two, or three protrusions, respectively . Thin-section and freeze-fracture electron microscopy of the protrusions showed a small triglyceride droplet surrounded by a thin cytoplasmic rim that was continuous with the main cytoplasmic matrix . The glycocalyx coating and plasma membrane extended from the cell surface onto, and over, the protrusion . Scanning microscopy of cells stimulated by lipolytic agents, including epinephrine, adrenocorticotropic hormone, theophylline, and dibutyryl cyclic AMP, revealed a dosedependent increase in the number of protrusions per cell hemisphere . Maximal concentrations of lipolytic hormones caused an average 2 .5-fold increase in the number of protrusions per hemisphere without changing the average size of the protrusions . Only 40% of the stimulated cell hemispheres exhibited no protrusions ; over 15% of the cells contained three or more ; and a number of the protrusions were multilobulate. Insulin prevented the increase in the number of protrusions and the change in distribution caused by the lipolytic hormones but did not prevent the increase caused by theophylline and dibutyryl cyclic AMP . The data suggest that the protrusions are a structural feature of the cell and may be related to the lipolytic pathway. These observations may help explain some of the discrepant biochemical data relating to hormonal stimulation of lipolysis .

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Volume changes during preparation of mouse embryonic tissue for scanning electron microscopy

1979

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A new approach to the study of cell organelles with the electron microscope

Cited by 12 publications

References 4 publications

Cell and organelle shrinkage during preparation for scanning electron microscopy: effects of fixation, dehydration and critical point drying

Cell and organelle shrinkage during preparation for scanning electron microscopy: effects of fixation, dehydration and critical point drying

Surface structure changes of rat adipocytes during lypolysis stimulated by various lypolysis stimulated by various lypolytic agents

Volume changes during preparation of mouse embryonic tissue for scanning electron microscopy

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