Abstract

Spectral curves of direct transmittance, diffuse transmittance, and scattering by partial layers of oriented red blood cells were measured. Also determined were the average optical parameters of a single cell and the cell concentration in the partial layers. The experiments are an extension and refinement of those of Lothian and Lewis on red blood cells and of Latimer on the green alga Chlorella. By assuming large-particle optics, equations were derived to predict the spectral curves in terms of cell and layer parameters. Experimental and theoretical curves are in reasonable agreement.

© 1961 Optical Society of America

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References

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  1. E. E. Jacobs, A. S. Holt, R. Kromhout, and E. Rabinowitch, Arch. Biochem. Biophys. 72, 495 (1957). See also E. Rabinowitch, Photosynthesis and Related Processes (Interscience Publishers, Inc., New York, 1956), Vol. II, Part 2, p. 1816.
    [CrossRef] [PubMed]
  2. S. Anderson, J. Opt. Soc. Am. 39, 49 (1949).
    [CrossRef] [PubMed]
  3. J. W. Weigl, J. Chem. Phys. 24, 577 (1956).
    [CrossRef]
  4. P. Latimer and E. Rabinowitch, Arch. Biochem. Biophys. 84, 428 (1959).
    [CrossRef] [PubMed]
  5. P. Latimer, Plant Physiol. 34, 193 (1959).See also, P. Latimer, Science 127, 29 (1958).
    [CrossRef] [PubMed]
  6. F. S. Brackett and E. Charney, J. Opt. Soc. Am. 50, 811 (1960). See also E. Charney and F. Brackett, Arch. Biochem. Biophys. 92, 1 (1961).
    [CrossRef] [PubMed]
  7. G. F. Lothian and P. C. Lewis, Nature 178, 1342 (1956). See also G. F. Lothian, Absorption Spectrophotometry (The MacMillan Company, New York, 1958), p. 35.
    [CrossRef]
  8. P. Latimer, thesis, University of Illinois, Urbana, Illinois (1956).
  9. K. Shibata, A. A. Benson, and M. Calvin, Biochim. et Biophys. Acta 15, 461 (1954).
    [CrossRef]
  10. S. Keilin and E. F. Hartree, Biochim. et Biophys. Acta 27, 173 (1958).
    [CrossRef]
  11. M. Born and E. Wolf, Principles of Optics (Pergamon Press, New York, 1959), p. 395.
  12. H. C. van de Hulst, Light Scattering by Small Particles (John Wiley & Sons, Inc., New York, 1957), p. 107 and 193.
  13. P. Latimer, J. Opt. Soc. Am. 51, 116 (1961).
    [CrossRef]
  14. M. M. Wintrobe, Clinical Hematology (Lea & Febiger Publishers, Philadelphia, Pennsylvania, 1951), 3rd ed., p. 312.
  15. R. Barer, Physical Techniques in Biological Research, III, edited by G. Oster and A. W. Pollister (Academic Press, Inc., New York, 1956), p. 29.
  16. D. L. Drabkin and J. H. Austin, J. Biol. Chem. 112, 51 (1951).
  17. R. Barer and S. Joseph, Quart. J. Microscop. Sci. 95, 399 (1954).

1961 (1)

1960 (1)

1959 (2)

P. Latimer and E. Rabinowitch, Arch. Biochem. Biophys. 84, 428 (1959).
[CrossRef] [PubMed]

P. Latimer, Plant Physiol. 34, 193 (1959).See also, P. Latimer, Science 127, 29 (1958).
[CrossRef] [PubMed]

1958 (1)

S. Keilin and E. F. Hartree, Biochim. et Biophys. Acta 27, 173 (1958).
[CrossRef]

1957 (1)

E. E. Jacobs, A. S. Holt, R. Kromhout, and E. Rabinowitch, Arch. Biochem. Biophys. 72, 495 (1957). See also E. Rabinowitch, Photosynthesis and Related Processes (Interscience Publishers, Inc., New York, 1956), Vol. II, Part 2, p. 1816.
[CrossRef] [PubMed]

1956 (2)

G. F. Lothian and P. C. Lewis, Nature 178, 1342 (1956). See also G. F. Lothian, Absorption Spectrophotometry (The MacMillan Company, New York, 1958), p. 35.
[CrossRef]

J. W. Weigl, J. Chem. Phys. 24, 577 (1956).
[CrossRef]

1954 (2)

K. Shibata, A. A. Benson, and M. Calvin, Biochim. et Biophys. Acta 15, 461 (1954).
[CrossRef]

R. Barer and S. Joseph, Quart. J. Microscop. Sci. 95, 399 (1954).

1951 (1)

D. L. Drabkin and J. H. Austin, J. Biol. Chem. 112, 51 (1951).

1949 (1)

Anderson, S.

Austin, J. H.

D. L. Drabkin and J. H. Austin, J. Biol. Chem. 112, 51 (1951).

Barer, R.

R. Barer and S. Joseph, Quart. J. Microscop. Sci. 95, 399 (1954).

R. Barer, Physical Techniques in Biological Research, III, edited by G. Oster and A. W. Pollister (Academic Press, Inc., New York, 1956), p. 29.

Benson, A. A.

K. Shibata, A. A. Benson, and M. Calvin, Biochim. et Biophys. Acta 15, 461 (1954).
[CrossRef]

Born, M.

M. Born and E. Wolf, Principles of Optics (Pergamon Press, New York, 1959), p. 395.

Brackett, F. S.

Calvin, M.

K. Shibata, A. A. Benson, and M. Calvin, Biochim. et Biophys. Acta 15, 461 (1954).
[CrossRef]

Charney, E.

Drabkin, D. L.

D. L. Drabkin and J. H. Austin, J. Biol. Chem. 112, 51 (1951).

Hartree, E. F.

S. Keilin and E. F. Hartree, Biochim. et Biophys. Acta 27, 173 (1958).
[CrossRef]

Holt, A. S.

E. E. Jacobs, A. S. Holt, R. Kromhout, and E. Rabinowitch, Arch. Biochem. Biophys. 72, 495 (1957). See also E. Rabinowitch, Photosynthesis and Related Processes (Interscience Publishers, Inc., New York, 1956), Vol. II, Part 2, p. 1816.
[CrossRef] [PubMed]

Jacobs, E. E.

E. E. Jacobs, A. S. Holt, R. Kromhout, and E. Rabinowitch, Arch. Biochem. Biophys. 72, 495 (1957). See also E. Rabinowitch, Photosynthesis and Related Processes (Interscience Publishers, Inc., New York, 1956), Vol. II, Part 2, p. 1816.
[CrossRef] [PubMed]

Joseph, S.

R. Barer and S. Joseph, Quart. J. Microscop. Sci. 95, 399 (1954).

Keilin, S.

S. Keilin and E. F. Hartree, Biochim. et Biophys. Acta 27, 173 (1958).
[CrossRef]

Kromhout, R.

E. E. Jacobs, A. S. Holt, R. Kromhout, and E. Rabinowitch, Arch. Biochem. Biophys. 72, 495 (1957). See also E. Rabinowitch, Photosynthesis and Related Processes (Interscience Publishers, Inc., New York, 1956), Vol. II, Part 2, p. 1816.
[CrossRef] [PubMed]

Latimer, P.

P. Latimer, J. Opt. Soc. Am. 51, 116 (1961).
[CrossRef]

P. Latimer and E. Rabinowitch, Arch. Biochem. Biophys. 84, 428 (1959).
[CrossRef] [PubMed]

P. Latimer, Plant Physiol. 34, 193 (1959).See also, P. Latimer, Science 127, 29 (1958).
[CrossRef] [PubMed]

P. Latimer, thesis, University of Illinois, Urbana, Illinois (1956).

Lewis, P. C.

G. F. Lothian and P. C. Lewis, Nature 178, 1342 (1956). See also G. F. Lothian, Absorption Spectrophotometry (The MacMillan Company, New York, 1958), p. 35.
[CrossRef]

Lothian, G. F.

G. F. Lothian and P. C. Lewis, Nature 178, 1342 (1956). See also G. F. Lothian, Absorption Spectrophotometry (The MacMillan Company, New York, 1958), p. 35.
[CrossRef]

Rabinowitch, E.

P. Latimer and E. Rabinowitch, Arch. Biochem. Biophys. 84, 428 (1959).
[CrossRef] [PubMed]

E. E. Jacobs, A. S. Holt, R. Kromhout, and E. Rabinowitch, Arch. Biochem. Biophys. 72, 495 (1957). See also E. Rabinowitch, Photosynthesis and Related Processes (Interscience Publishers, Inc., New York, 1956), Vol. II, Part 2, p. 1816.
[CrossRef] [PubMed]

Shibata, K.

K. Shibata, A. A. Benson, and M. Calvin, Biochim. et Biophys. Acta 15, 461 (1954).
[CrossRef]

van de Hulst, H. C.

H. C. van de Hulst, Light Scattering by Small Particles (John Wiley & Sons, Inc., New York, 1957), p. 107 and 193.

Weigl, J. W.

J. W. Weigl, J. Chem. Phys. 24, 577 (1956).
[CrossRef]

Wintrobe, M. M.

M. M. Wintrobe, Clinical Hematology (Lea & Febiger Publishers, Philadelphia, Pennsylvania, 1951), 3rd ed., p. 312.

Wolf, E.

M. Born and E. Wolf, Principles of Optics (Pergamon Press, New York, 1959), p. 395.

Arch. Biochem. Biophys. (2)

P. Latimer and E. Rabinowitch, Arch. Biochem. Biophys. 84, 428 (1959).
[CrossRef] [PubMed]

E. E. Jacobs, A. S. Holt, R. Kromhout, and E. Rabinowitch, Arch. Biochem. Biophys. 72, 495 (1957). See also E. Rabinowitch, Photosynthesis and Related Processes (Interscience Publishers, Inc., New York, 1956), Vol. II, Part 2, p. 1816.
[CrossRef] [PubMed]

Biochim. et Biophys. Acta (2)

K. Shibata, A. A. Benson, and M. Calvin, Biochim. et Biophys. Acta 15, 461 (1954).
[CrossRef]

S. Keilin and E. F. Hartree, Biochim. et Biophys. Acta 27, 173 (1958).
[CrossRef]

J. Biol. Chem. (1)

D. L. Drabkin and J. H. Austin, J. Biol. Chem. 112, 51 (1951).

J. Chem. Phys. (1)

J. W. Weigl, J. Chem. Phys. 24, 577 (1956).
[CrossRef]

J. Opt. Soc. Am. (3)

Nature (1)

G. F. Lothian and P. C. Lewis, Nature 178, 1342 (1956). See also G. F. Lothian, Absorption Spectrophotometry (The MacMillan Company, New York, 1958), p. 35.
[CrossRef]

Plant Physiol. (1)

P. Latimer, Plant Physiol. 34, 193 (1959).See also, P. Latimer, Science 127, 29 (1958).
[CrossRef] [PubMed]

Quart. J. Microscop. Sci. (1)

R. Barer and S. Joseph, Quart. J. Microscop. Sci. 95, 399 (1954).

Other (5)

P. Latimer, thesis, University of Illinois, Urbana, Illinois (1956).

M. Born and E. Wolf, Principles of Optics (Pergamon Press, New York, 1959), p. 395.

H. C. van de Hulst, Light Scattering by Small Particles (John Wiley & Sons, Inc., New York, 1957), p. 107 and 193.

M. M. Wintrobe, Clinical Hematology (Lea & Febiger Publishers, Philadelphia, Pennsylvania, 1951), 3rd ed., p. 312.

R. Barer, Physical Techniques in Biological Research, III, edited by G. Oster and A. W. Pollister (Academic Press, Inc., New York, 1956), p. 29.

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Figures (6)

Fig. 1
Fig. 1

Schematic diagram of relation of the partial layer of red blood cells to the light intensities observed. To measure (∑It+∑Is)/∑I0, an opal glass diffusing plate was placed on top of both the sample and the blank vessels.

Fig. 2
Fig. 2

Parameters of red blood cells used in calculations.

Fig. 3
Fig. 3

Extinction (log 1/direct transmittance) curves and absorption curve of aging red blood cells with attempts to explain results theoretically by Eq. (8) with ρ=0. The constants on the ordinate scale were selected for convenience in plotting. The cell thickness for the theoretical absorption curve was chosen to give Q=1.

Fig. 4
Fig. 4

Extinction curves of different layer concentrations of red blood cells and absorption curve of one such layer. The theoretical extinction curves were calculated from Eq. (9) and the absorption curve from Eq. (3), all with ρ corrections.

Fig. 5
Fig. 5

Small-angle scattering by red blood cells. The ordinate scale gives absolute ratios if the constant is 224 for 1.5° and 586 for 3.3°.

Fig. 6
Fig. 6

Direct transmittance, total small-angle scattering, and the sum of absorptance and reflectance of a partial layer of red blood cells (Q=0.37, t=1.81).

Equations (10)

Equations on this page are rendered with MathJax. Learn more.

I = I 0 10 - α c L ,
( Σ I t + Σ I s ) / Σ I 0 = ( 1 - Q ) + Q 10 - α c t ,
( Σ I t + Σ I s ) / Σ I 0 = ( 1 - Q ) + 1 2 Q 10 - α c t * + 1 2 Q 10 - 2 α c t * .
( Σ I t + Σ I s ) / Σ I 0 = ( 1 - Q ) + Q 10 - ( α c t + ρ ) .
a d ( 0 ) = k d A 0 10 - 1 2 α c L ,
a a ( 0 ) = k a A 0 10 - α c t * .
a s ( 0 ) = k s A 0 .
I t / I 0 = ( 1 - Q ) 2 + Q 2 10 - ( α c t + ρ ) + 2 Q ( 1 - Q ) 10 - 1 2 ( α c t + ρ ) cos ϕ
I t / I 0 = [ ( 1 - Q ) + 1 2 Q ( 10 - 1 2 ( α c t * + ρ ) cos ϕ * + 10 - 1 2 ( 2 α c t * + ρ ) cos 2 ϕ * ) ] 2 + [ 1 2 Q ( 10 - 1 2 ( α c t * + ρ ) × sin ϕ * + 10 - 1 2 ( 2 α c t * + ρ ) sin 2 ϕ * ) ] 2 .
Σ I s Σ I 0 = Σ I t + Σ I s Σ I 0 - I t I 0 .