Skip to main content

Light Scattering Properties of Murine Hemopoietic Cells

  • Conference paper
Book cover Automation in Hematology

Abstract

The light scatter of cells contains information about cell structure and size. Much of this information can be obtained by measuring the light scattered in a flow cytometer in two directions: forward (1.5°–13°) and perpendicular (65°–115°) with respect to the direction of the laser light. For different mouse bone marrow cell types and Sephadex G-25 beads of 10–50 μm diameter, the forward light scatter intensity can be shown to be linearly proportional to the cross-sectional area. The perpendicular light scatter intensity can be shown to depend both on size and degree of structuredness. Therefore, light scatter measurements may be used to obtain overall morphological descriptions of rare cells. By sorting on the basis of light scatter measurements and by subsequent in vivo and in vitro culture assays, it can be shown that the pluripotent hemopoietic stem cell and three committed progenitor cells which represent consecutive stages in the granulocyte/monocyte differentiation series have diameters of 7.1–7.5 μm, and show a complexity of structuredness which increases with differentiation. Since these cells have a low incidence and are only described by their function, such morphological information cannot be obtained by direct microscopic examination of bone marrow.

Furthermore, most measurements by flow cytometers can be improved by simultaneous light scatter measurements. Examples are presented which illustrate this in studies of immunofluorescence, leukemic bone marrow, and stem cell purification.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Chapter
USD 29.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 109.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. ARNDT-JOVIN, D. J., JOVIN, T. M.: Computer-controlled multiparameter analysis and sorting of cells and particles. J. Histochem. Cytochem. 22, 622–625, 1974

    Article  CAS  PubMed  Google Scholar 

  2. ARNDT-JOVIN, D. J., JOVIN, T. M.: Analysis and sorting of living cells according to deoxyribonucleic acid content. J. Hisochem. Cytochem. 25, 585–589, 1977

    Article  CAS  Google Scholar 

  3. BEKKUM, D. W. VAN, NOORD, M. J. VAN, MAAT, A., DICKE, K. A.: Attempts at identification of hemopoietic stem cell in mouse. Blood 38, 547–558, 1971

    PubMed  Google Scholar 

  4. BEKKUM, D. W., VAN: The appearance of the multipotential hemopoietic stem cell. In: BAUM, S. J., LEDNEY, G. D., eds., Experimental Hematology Today. Berlin, Heidelberg, New York, Verlag Springer 1977, pp. 3–10

    Chapter  Google Scholar 

  5. BOL, S., ENGH, G. VAN DEN, VISSER, J.: A technique for staining haemopoietic colonies in agar cultures. Exp. Hematol. 5, 551–553, 1977

    CAS  PubMed  Google Scholar 

  6. BOL, S., VISSER, J.W.M., WILLIAMS, N., ENGH, G. VAN DEN: Physical characterization of haemopoietic progenitor cells by equilibrium density centrifugation. In: BLOEMENDAL, H., ed., Cell Separation Methods. Amsterdam, Elsevier/North-Holland, 1977, pp. 41–52

    Google Scholar 

  7. BOL, S.: The detection of three different states of the monocyte/granulocyte progenitor cell from normal mouse bone marrow. IN: BENTVELZEN, P., HILGERS, J., YOHN, D. S., eds., Advances in Comparative Leukemia Research. Amsterdam, Elsevier/North-Holland Biomedical Press, 1978, pp. 283–284

    Google Scholar 

  8. BOL, S., VISSER, J., ENGH, G. VAN DEN: The physical separation of three subpopulations of granulocyte/macrophage progenitor cells from mouse bone marrow. Exp. Hematol. 7, 541–553, 1979

    CAS  PubMed  Google Scholar 

  9. CICCO, D. DE, GREAVES, M. F., ENGH, G. VAN DEN, WARNER, N. L.: Expression of choleratoxin binding sites on normal and malignant hemopoietic cells. In: BENTVELZEN, P., HILGERS, J., YOHN, D. S., eds., Advances in Comparative Leukemia Research. Amsterdam, Elsevier/North-Holland Biomedical Press, 1978, pp. 357–362

    Google Scholar 

  10. HAAIJMAN, J. J., DALEN, J. P. R. VAN: Quantification in immunofluorescence microscopy a new standard for fluorescein and rhodamine emission measurement. J. Immunol. Methods 5, 359–374, 1974

    Article  CAS  PubMed  Google Scholar 

  11. HERZENBERG, L. A., SWEET, R. G., HERZENBERG, L. A.: Fluorescence-activated cell sorting. Sci. Am. 234, 108–117, 1976

    Article  CAS  PubMed  Google Scholar 

  12. HULST, H. C. VAN DE: Light Scattering by Small Particles. New York, Wiley 1957

    Google Scholar 

  13. JOVIN, T. M., MORRIS, S. J., STRIKER, G., SCHULTENS, H. A., DIGWEED, M., ARNDT-JOVIN, D. J.: Automatic sizing and separation of particles by ratios of light scattering intensities. J. Histochem. Cytochem. 24, 269–283, 1976

    Article  CAS  PubMed  Google Scholar 

  14. KACHEL, V., KORDWIG, E., GLOSSNER, E.: Uniform lateral orientation, caused by flow forces, of flat particles in flow-through systems. J. Histochem. Cytochem. 25, 774–780, 1977

    Article  CAS  PubMed  Google Scholar 

  15. KERKER, M.: The Scattering of Light and Other Electromagnetic Radiation. New York, Academic Press, 1969

    Google Scholar 

  16. KERKER, M., CHEW, H., MULTHY, P. J., KRATOHVIL, J. P., COOKE, D. D., SCULLEY, M., LEE, M. P.: Light scattering and fluorescence by small particles having internal structure. J. Histochem. Cytochem. 27, 250–263, 1979

    Article  CAS  PubMed  Google Scholar 

  17. LATT, S. A., GEORGE, Y. S., GRAY, J. W.: Flow cytometric analysis of bromodeoxyuridine substituted cells stained with 33258 Hoechst. J. Histochem. Cytochem. 25, 927–934, 1977

    Article  CAS  PubMed  Google Scholar 

  18. LESSIN, L. S., BESSIS, M.: Morphology of the erythron. In: WILLIAMS, W. J., BEUTLER, E., ERSLEV, A. J., RUNDLES, R. W., eds., Hematology. New York, McGraw-Hill, 1972, pp. 62–93

    Google Scholar 

  19. LOKEN, M. R., SWEET, R. G., HERZENBERG, L. A.: Cell discrimination by multiangle light scattering. J. Histochem. Cytochem. 24, 284–291, 1976

    Article  CAS  PubMed  Google Scholar 

  20. LOKEN, M. R., PARKS, D. R., HERZENBERG, L. A.: Identification of cell asymmetry and orientation by light scattering. /. Histochem. Cytochem. 25, 790–795, 1977

    Article  CAS  Google Scholar 

  21. MARIKOVSKY, Y., KHODADAD, J. K., WEINSTEIN, R. S.: Influence of red cell shape on surface charge topography. Exp. Cell Res. 116, 191–197, 1978

    Article  CAS  PubMed  Google Scholar 

  22. MEYER, R. A., BRUNSTING, A.: Light scattering from nucleated biological cells. Biophys. J. 15, 191–203, 1975

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  23. MULLANEY, P. F., VAN DILLA, M. A., COULTER, J. R., DEAN, P. N.: Cell sizing: A light scattering photometer for rapid volume determination. Rev. Sci. Instrum. 40, 1029–1032 1969

    Article  CAS  PubMed  Google Scholar 

  24. ROEDER, C.: Surface charge and sialic acid content of L5222 and BNML leukemia cells. Leuk. Res. 1, 201–205, 1977

    Article  CAS  Google Scholar 

  25. SALZMAN, G. C., CROWELL, J. M., MARTIN, J. C., TRUJILLO, T. T., ROMERO, A., MULLANEY, P. F., LA BAUVE, P. M.: Cell classification by laser light scattering: Identification and separation of unstained leukocytes. Acta Cytol. 19, 374–377, 1975

    CAS  PubMed  Google Scholar 

  26. SALZMAN, G. C., PRICE, B. J., HIEBERT, R. D., MULLANEY, F., VISSER, J. W. M.: Laser back scattering measurements on biological cells: Preliminary results. In: BLOEMENDAL, H., ed., Cell Separation Methods. Amsterdam, Elsevier/North-Holland Biomedical Press, 1977, pp. 67–78

    Google Scholar 

  27. SALZMAN, G. C., WILDER, M. E., JETT, J. H.: Light scattering with stream-in-air flow systems. J. Histochem. Cytochem. 27, 264–267, 1979

    Article  CAS  PubMed  Google Scholar 

  28. SHARPLESS, T. K., BARTHOLDI, M., MELAMED, M. R.: Size and refractive index dependence of simple forward angle scattering measurements in a flow system using sharply focussed illumination. J. Histochem. Cytochem. 25, 845–856, 1977

    Article  CAS  PubMed  Google Scholar 

  29. TILL, J. E., MCCULLOCH, E. A.: Direct measurements of radiation sensitivity of normal bone marrow cells. Radiat. Res. 14, 213–220, 1961

    Article  CAS  PubMed  Google Scholar 

  30. VISSER, J., ENGH, G. J. VAN DEN, WILLIAMS, N., MULDER, D.: Physical separation of the cycling and noncycling compartments of murine hemopoietic stem cells. IN: BAUM, S. J., LEDNEY, G. D., eds., Experimental Hematology Today. New York, Springer, 1979, pp. 21–27

    Google Scholar 

  31. VISSER, J. W. M., CRAM, L. S., MARTIN, J. C., SALZMAN, G. C., PRICE, B. J.: Sorting of a murine granulocytic progenitor cell by use of laser light scattering measurements. In: LUTZ, D., ed., Pulse-Cytophotometry, Part III. Ghent, European Press, 1978, pp. 187–192

    Google Scholar 

  32. VISSER, J., ENGH, G. J. VAN DEN, WILLIAMS, N., MULDER, D.: Buoyant densities and sedimentation rates of the murine haemopoietic stem cells. In: BENTVELZEN, P., HILGERS, J., YOHN, D. S., eds., Advances in Comparative Leukemia Research. Amsterdam, Elsevier/North-Holland Biomedical Press, 1978, pp. 279–282

    Google Scholar 

  33. VISSER, J., HAAIJMAN, J., TRASK, B.: Quantitative immunofluorescence in flow cytometry. In: KNAPP, W., HOLUBAR, K., WICK, G., eds., Immunofluorescence and Related Staining Techniques. Amsterdam, Elsevier/North-Holland Biomedical Press, 1978, pp. 147–160

    Google Scholar 

  34. WAGEMAKER, G., OBER-KIEFTENBERG, V. E., BROUWER, A., PETERS-SLOUGH, M. F.: Some characteristics of in vitro erythroid colony and burst forming units. In: BAUM, S. J., LEDNEY, G. D., eds., Experimental Hematology Today. Berlin, Heidelberg, New York, Springer Verlag, 1977, pp. 103–110

    Chapter  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 1981 Springer-Verlag Berlin Heidelberg

About this paper

Cite this paper

Visser, J.W.M., van den Engh, G.J., van Bekkum, D.W. (1981). Light Scattering Properties of Murine Hemopoietic Cells. In: Ross, D.W., Brecher, G., Bessis, M. (eds) Automation in Hematology. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-67756-4_22

Download citation

  • DOI: https://doi.org/10.1007/978-3-642-67756-4_22

  • Publisher Name: Springer, Berlin, Heidelberg

  • Print ISBN: 978-3-540-10225-0

  • Online ISBN: 978-3-642-67756-4

  • eBook Packages: Springer Book Archive

Publish with us

Policies and ethics