Structural, Biochemical, and Elemental Correlates of Injury in Cultured Cardiac Cells

LeFurgey, Ann; Murphy, Elizabeth; Wagenknecht, Bernhard; Ingram, Peter; Lieberman, Melvyn

doi:10.1007/978-1-4684-5598-4_59

Cell Calcium Metabolism 1989

DOI: 10.1007/978-1-4684-5598-4_59

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Structural, Biochemical, and Elemental Correlates of Injury in Cultured Cardiac Cells

Ann LeFurgey

Elizabeth Murphy

Bernhard Wagenknecht

et al.

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1992

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“…Monolayers of cardiac cells also have been frozen by freeze-clamping methods (Buja et al, 1985;Jones e t al., 1989;Morris et al, 1989). The aim of the present investigation was to develop an in situ, quick-freezing technique for chicken cardiac cell cultures, which in our laboratory serve as an experimental model both for studying membrane ion transport (Horres et al, 1987), and for studying the influence of anoxia and substrate deprivation on the cellular metabolism and ultrastructure (Murphy et al, 1987;LeFurgey et al, 1989).…”

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Quick‐freezing of cultured cardiac cells in situ with special attention to the mitochondrial ultrastructure

Dalen

Lieberman

LeFurgey

et al. 1992

Journal of Microscopy

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SUMMARY A new method has been developed which allows quick‐freezing in situ of primary, cardiac cell cultures grown to confluence on gas‐permeable membranes (Petriperm dishes). Small pieces of the growth substratum, with rhythmically beating myocardial cells, were slam‐frozen, without cryoprotectants, against the surface of a helium‐cooled copper block at approximately 16 K. The quality of the cellular cryopreservation, as judged by ultrastructural criteria, was studied in freeze‐substituted specimens processed for transmission electron microscopy. The ultrastructure of cryofixed cardiac cells was compared with that of unfrozen, chemically fixed samples. The severity of cryodistortions increased progressively with increasing distance from the point of first impact. Of particular interest were the dramatic alterations of the mitochondrial ultrastructure. The concept that the reticular and the outer mitochondrial membranes are intimately and strongly associated was clearly demonstrated. Optimally frozen material revealed cryopreserved ultrastructure of high quality. The method described appears to offer an ideal model system for correlating the information gained by phase‐contrast microscopy of living cell cultures with the ultrastructure of the same samples fixed in situ by chemical or physical techniques. Cryofixation would be particularly useful for studying dynamic cellular processes associated with physiological and pathophysiological conditions, e.g. metabolic inhibition, anoxia and substrate deprivation.

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mentioning

confidence: 99%

Quick‐freezing of cultured cardiac cells in situ with special attention to the mitochondrial ultrastructure

Dalen

Lieberman

LeFurgey

et al. 1992

Journal of Microscopy

Self Cite

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Structural, Biochemical, and Elemental Correlates of Injury in Cultured Cardiac Cells

Cited by 1 publication

References 14 publications

Quick‐freezing of cultured cardiac cells in situ with special attention to the mitochondrial ultrastructure

Quick‐freezing of cultured cardiac cells in situ with special attention to the mitochondrial ultrastructure

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