Structural artifacts and advantages of cytocentrifugation of cells as viewed by scanning electron microscopy.
Cytocentrifugation of cell suspensions onto glass slides is a widely used procedure in contemporary cytology. We employed here scanning electron microscopy (SEM) to investigate putative morphological changes induced in cells submitted to cytocentrifugation. The fine structure of murine pleural exudate cells (macrophages mainly) processed by spinning was compared with that of similar cells treated without centrifugation (poly-L-lysine attachment of the cells to glass slides at 1 g). Cells of cytocentrifuged preparations showed a significant increase in diameter and smoothening of the cell surface as compared with the morphology of non-centrifuged cells. Cytocentrifugation also induced the formation of thin elongations coming out of the cellular outlines. The centrifugation-induced flattening of the pleural macrophages improved the detection of large intracellular inclusions (containing tungsten particles): these bodies were readily identified by secondary-electron imaging mode of SEM in cytospinned cells whereas their detection in non-centrifuged spherical cells required the use of the backscattered-electron imaging mode of SEM. We conclude that the cytocentrifugation methodology, on one hand, requires caution on the interpretation of the microanatomy of the cells and, on the other hand, the procedure may be an adequate method to improve the identification of large intracellular inclusions by routine (secondary-electron imaging mode) SEM.
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Depart. of Anatomy, Abel Salazar Institute for the Biomedical Sciences, University of Porto, Portugal.
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de Sá
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Grande
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D000818:Animals; D016477:Artifacts; D002498:Centrifugation / Q000379:methods*; D005260:Female; D051379:Mice; D008855:Microscopy, Electron, Scanning / Q000379:methods*; D010994:Pleura / Q000166:cytology / Q000648:ultrastructure