OSA's Digital Library

Applied Optics

Applied Optics


  • Vol. 24, Iss. 9 — May. 1, 1985
  • pp: 1355–1365

Flow-cytometric light scattering measurement of red blood cell volume and hemoglobin concentration

D. H. Tycko, M. H. Metz, E. A. Epstein, and A. Grinbaum  »View Author Affiliations

Applied Optics, Vol. 24, Issue 9, pp. 1355-1365 (1985)

View Full Text Article

Enhanced HTML    Acrobat PDF (1372 KB)

Browse Journals / Lookup Meetings

Browse by Journal and Year


Lookup Conference Papers

Close Browse Journals / Lookup Meetings

Article Tools



A light scattering technique for simultaneously determining the volume (V) and hemoglobin concentration (HC) of individual sphered red blood cells (RBCs) is described. Light scattered into two angular intervals yields measurements S1 and S2, respectively. Since a sphered RBC is essentially a homogeneous dielectric sphere having a complex refractive index that is linear in HC, with a proper choice of detector acceptance angles, tables relating V and HC to S1 and S2 can be computed via Mie theory. Absolute calibration is possible using droplets of water-immisible oils of accurately known refractive index. Results of experimental tests of the method are compared with those obtained from hematological reference measurements.

© 1985 Optical Society of America

Original Manuscript: October 19, 1984
Published: May 1, 1985

D. H. Tycko, M. H. Metz, E. A. Epstein, and A. Grinbaum, "Flow-cytometric light scattering measurement of red blood cell volume and hemoglobin concentration," Appl. Opt. 24, 1355-1365 (1985)

Sort:  Author  |  Year  |  Journal  |  Reset  


  1. For concise reviews of the subject and extensive bibliographies, see T. M. Jovin et al., “Automatic Sizing and Separation of Particles by Ratios of Light Scattering Intensities,” J. Histochem. Cytochem. 24, 269 (1976);G. C. Salzman, “Flow Cytometry: The Use of Lasers for Rapid Analysis and Separation of Single Biological Cells,” in The Biomedical Laser, L. Goldman, Ed. (Springer, New York, 1981), Chap. 5. [CrossRef] [PubMed]
  2. L. P. Bayvel, A. R. Jones, Electromagnetic Scattering and Its Applications (Applied Science, London, 1981), pp. 22–47.
  3. M. M. Wintrobe et al., Clinical Hematology (Lea & Febiger, Philadelphia, 1981), p. 18.
  4. J. D. Bessman, R. K. Johnson, “Erythrocyte Volume Distribution in Normal and Abnormal Subjects,” Blood 46, 369 (1975). [PubMed]
  5. H. E. Kubitschek, “Electronic Measurement of Particle Size,” Res. Appl. Ind. 13, 128 (1960).
  6. N. Mohandas et al., “Inaccuracies Associated with the Automated Measurement of MCHC in Dehydrated Cells,” Blood 56, 125 (1980). [PubMed]
  7. T. Arnfred, S. D. Kristensen, V. Munck, “Colter Counter Model S and Model S-Plus Measurements of Mean Erythrocyte Volume (MCV) are Influenced by the Mean Erythrocyte Haemoglobin Concentration (MCHC),” Scand. J. Clin. Lab. Invest. 41, 717 (1981). [CrossRef]
  8. Y. R. Kim, L. Ornstein, “Isovolumetric Sphering of Erythrocyte for More Accurate and Precise Cell Volume Measurement by Flow Cytometry,” Cytometry 3, 419 (1983). [CrossRef] [PubMed]
  9. M. Kerker, The Scattering of Light and Other Electromagnetic Radiation (Academic, New York, 1969), Chap. 3.
  10. Ref. 3, Chap. 4.
  11. R. Barer, S. Joseph, “Refractometry of Living Cells,” Q. J. Microsc. Sci. 95, 399 (1954).
  12. Ref. 9, chap. 8.
  13. R. B. Pennell, “Composition of Normal Human Red Cells,” in The Red Blood Cell, C. Bishop, D. M. Surgenor, Eds. (Academic, New York, 1964), Chap. 2.
  14. R. S. Longhurst, Geometrical and Physical Optics (Longman, London, 1967), pp. 446–448.
  15. O. W. van Assendelft, Spectrophotometry of Haemoglobin Derivatives (van Gorcum, Assen, The Netherlands, 1970, pp. 23–25 and Chap. 3 and Chap. 3.
  16. H. P. Mansberg, A. M. Saunders, W. Groner, “The Hemalog D White Cell Differential System,” J. Histochem. Cytochem. 22, 711 (1974). [CrossRef] [PubMed]
  17. National Committee for Clinical Laboratory Standards (NCCLS), Publication H7-T (1981).
  18. International Committee for Standardization in Haematology, J. Clin. Pathol. 31, 139 (1978). [PubMed]
  19. M. R. Clark et al., “Study on the Dehydrating Effect of the Red Cell Na+/K+-Pump in Nystatin-Treated Cells with Varying Na+ and Water Contents,” Biochim. Biophys. Acta 646, 422 (1981). [CrossRef] [PubMed]
  20. L. M. Corash et al., “Separation of Erythrocytes According to Age on a Simplified Density Gradient,” J. Lab. Clin. Med. 84, 147 (1974). [PubMed]
  21. M. R. Clark, A. C. Greenquist, S. B. Shohet, “Stabilization of the Shape of Sickled Cells by Calcium and A23187,” Blood 48, 899 (1976). [PubMed]
  22. L. Ornstein, Mt. Sinai School of Medicine; private communication.

Cited By

Alert me when this paper is cited

OSA is able to provide readers links to articles that cite this paper by participating in CrossRef's Cited-By Linking service. CrossRef includes content from more than 3000 publishers and societies. In addition to listing OSA journal articles that cite this paper, citing articles from other participating publishers will also be listed.

« Previous Article  |  Next Article »

OSA is a member of CrossRef.

CrossCheck Deposited