Skip to main content
Biophysical Journal logoLink to Biophysical Journal
. 1987 Dec;52(6):947–954. doi: 10.1016/S0006-3495(87)83287-3

Differential polarization imaging. III. Theory confirmation. Patterns of polymerization of hemoglobin S in red blood sickle cells.

D A Beach 1, C Bustamante 1, K S Wells 1, K M Foucar 1
PMCID: PMC1330093  PMID: 3427201

Abstract

In this paper we test the predictions of the differential polarization imaging theory developed in the previous two papers. A characterization of the patterns of polymerization of hemoglobin in red blood cells from patients with sickle cell anemia is presented. This system was chosen because it is relatively easy to handle and because previous studies have been done on it. A differential polarization microscope designed and built in our laboratory was used to carry out this study. This microscope uses an image dissector camera, a photoelastic modulator, and a phase-lock amplifier. This design represents a substantial modification with respect to the instrumentation used in the previous results communicated on this system. Therefore, the results presented here also permit us to confirm the validity of our conclusions. On the basis of the differential polarization images obtained, models of the patterns of polymerization of the hemoglobin S inside the sickle cells are proposed and their M12 and regular images are calculated by the theory. Good agreement between those models and the experimental systems is found, as well as with the results previously reported.

Full text

PDF
951

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. ALLISON A. C. Properties of sickle-cell haemoglobin. Biochem J. 1957 Feb;65(2):212–219. doi: 10.1042/bj0650212. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bertles J. F., Döbler J. Reversible and irreversible sickling: a distinction by electron microscopy. Blood. 1969 Jun;33(6):884–898. [PubMed] [Google Scholar]
  3. Dykes G., Crepeau R. H., Edelstein S. J. Three-dimensional reconstruction of the fibres of sickle cell haemoglobin. Nature. 1978 Apr 6;272(5653):506–510. doi: 10.1038/272506a0. [DOI] [PubMed] [Google Scholar]
  4. Finch J. T., Perutz M. F., Bertles J. F., Döbler J. Structure of sickled erythrocytes and of sickle-cell hemoglobin fibers. Proc Natl Acad Sci U S A. 1973 Mar;70(3):718–722. doi: 10.1073/pnas.70.3.718. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Goldberg M. A., Lalos A. T., Bunn H. F. The effect of erythrocyte membrane preparations on the polymerization of sickle hemoglobin. J Biol Chem. 1981 Jan 10;256(1):193–197. [PubMed] [Google Scholar]
  6. Hofrichter J., Hendricker D. G., Eaton W. A. Structure of hemoglobin S fibers: optical determination of the molecular orientation in sickled erythrocytes. Proc Natl Acad Sci U S A. 1973 Dec;70(12):3604–3608. doi: 10.1073/pnas.70.12.3604. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Magdoff-Fairchild B., Rosen L. S. Plausible models of the sickle hemoglobin fiber based on x-ray diffraction data. Biophys J. 1986 Jan;49(1):67–69. doi: 10.1016/S0006-3495(86)83595-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Mickols W., Maestre M. F., Tinoco I., Jr, Embury S. H. Visualization of oriented hemoglobin S in individual erythrocytes by differential extinction of polarized light. Proc Natl Acad Sci U S A. 1985 Oct;82(19):6527–6531. doi: 10.1073/pnas.82.19.6527. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Mizukami H., Bartnicki D. E., Burke S., Brewer G. J., Mizukami I. F. The effect of erythrocyte membrane on the birefringence formation of sickle cell hemoglobin. Am J Hematol. 1986 Mar;21(3):233–241. doi: 10.1002/ajh.2830210302. [DOI] [PubMed] [Google Scholar]
  10. Peetermans J., Nishio I., Ohnishi S. T., Tanaka T. Light-scattering study of depolymerization kinetics of sickle hemoglobin polymers inside single erythrocytes. Proc Natl Acad Sci U S A. 1986 Jan;83(2):352–356. doi: 10.1073/pnas.83.2.352. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Ross P. D., Hofrichter J., Eaton W. A. Calorimetric and optical characterization of sickle cell hemoglobin gelation. J Mol Biol. 1975 Aug 5;96(2):239–253. doi: 10.1016/0022-2836(75)90345-9. [DOI] [PubMed] [Google Scholar]
  12. Shibata K., Cottam G. L., Waterman M. R. Acceleration of the rate of deoxyhemoglobin S polymerization by the erythrocyte membrane. FEBS Lett. 1980 Jan 28;110(1):107–110. doi: 10.1016/0014-5793(80)80034-2. [DOI] [PubMed] [Google Scholar]
  13. White J. G. Ultrastructural features of erythrocyte and hemoglobin sickling. Arch Intern Med. 1974 Apr;133(4):545–562. [PubMed] [Google Scholar]

Articles from Biophysical Journal are provided here courtesy of The Biophysical Society

RESOURCES