Cross-sectionalviewsofhemoglobinSfibersbyelectron.PDFVIP

Proc. Nati. Acad. Sci. USA Vol 76, No. 3, pp. 1140-1144, March 1979 Biochemistry Cross-sectional views of hemoglobin S fibers by electron microscopy and computer modeling (sickle cell hemoglobin fibers/thin sections/tannic acid embedding) ROBIN L. GARRELL, RICHARD H. CREPEAU, AND STUART J. EDELSTEIN Section of Biochemistry, Molecular and Cell Biology, Cornell University, Ithaca, New York 14853 Communicated by Helen M. Ranney, December 13,1978 ABSTRACT Fibers of deoxyHb S have been investigated but the possibility that distortions might arise during the em- by thin-section electron microscopy, utilizing a tannic acid bedding and sectioning of sickled cells cannot be completely embedding procedure. On the basis of numerous measurements excluded. of cross-sectional center-to-center distances for adjacent fibers Perhaps the most direct evidence that could be obtained on in + ± A pairs or arrays, fiber diameters (mean SD)of 205 5 in the issue of the fiber form present in sickled cells would be to embedded cells and 212 ± 8 A in embedded hemolysates were count of Hb molecules in cross sections of obtained. This is in agreement with values obtained by con- the strands S present ventional embedding procedures [Crepeau, R. H., Dykes, G., embedded fibers. In extensive studies using conventional em- Garrell, R. L. Edelstein, S. J. (1978) Nature (London)274, bedding techniques to determine the diameter of fibers in 616-617]. The use of tannic acid has resulted in improved res- sickled cells and cell-free preparations, however, the fiber cross olution of fiber cross sections, revealing individual strands of