Abstract

In order to assess the capability to optically identify small marine microbes, both simulations and experiments of angular resolved light scattering (ARLS) were performed. After calibration with 30-nm vesicles characterized by a nearly constant scattering distribution for vertically polarized light (azimuthal angle=90°), ARLS from suspensions of three types of marine picoplankton (two prokaryotes and one eukaryote) in seawater was measured with a scattering device that consisted of an elliptical mirror, a rotating aperture, and a PMT. Scattered light was recorded with adequate signal-to-noise in the 40–140°. Simulations modeled the cells as prolate spheroids with independently measured dimensions. For the prokaryotes, approximated as homogeneous spheroids, simulations were performed using the RM (Rayleigh-Mie) – I method, a hybrid of the Rayleigh-Debye approximation and the generalized Lorentz-Mie theory. For the picoeukaryote, an extended RM – I method was developed for a coated spheroid with different shell thickness distributions. The picoeukaryote was then modeled as a coated sphere with a spherical core. Good overall agreements were obtained between simulations and experiments. The distinctive scattering patterns of the different species hold promise for an identification system based on ARLS.

© 2006 Optical Society of America

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D. Watson, N. Hagen, J. Diver, P. Marchand, and M. Chachisvilis, "Elastic light scattering from single cells: orientational dynamics in optical trap," Biophys. J. 87, 1298-1306 (2004).
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A. Quirantes and S. Bernard, "Light scattering by marine algae: two-layer spherical and nonspherical models," J. Quant. Spectrosc. Radiat. Transfer 89,311-321 (2004).
[CrossRef]

2003

M. E. Lee and M. R. Lewis, "A New Method for the Measurement of the Optical Volume Scattering Function in the Upper Ocean," J. Atmos Ocean. Tech. 20, 563-571 (2003).
[CrossRef]

2002

Z. Ulanowski, R. S. Greenaway, P. H. Kaye, and I. K. Ludlow, "Laser diffractometer for single-particle scattering measurements," Meas. Sci. Technol 13, 292-296 (2002).
[CrossRef]

J. F. Kasting and J. L. Siefert, "Life and the evolution of Earth's atmosphere," Science 296, 1066-1068 (2002)
[CrossRef] [PubMed]

2001

2000

1999

1998

H. Volten, J. F. de Haan, J. W. Hovenier, R. Schreurs, W. Vassen, A. G. Dekker, H. J. Hoogenboom, F. Charlton, R. Wouts, "Laboratory measurements of Angular Distributions of light scattered by Phytoplankton and Silt," Limnol. Oceanogr. 43, 1180-1197 (1998).
[CrossRef]

1997

D. Marie, F. Partensky, S. Jacquet, and D. Vaulot, "Enumeration and cell cycle analysis of Natural Populations of Marine Picoplankton by Flow Cytometry using the nucleic acid stain SYBR Green I," Appl. Environ. Microbiol. 63, 186-193 (1997).
[PubMed]

1996

Eyvind Aas, "Refractive index of phytoplankton derived from its metabolite composition," J. Plankton Res. 18,2223-2249 (1996).
[CrossRef]

1991

R. C. MacDonald, R. I. MacDonald, B. P. M. Menco, K. Takeshita, N. K. Subbarao, and L.-r. Hu, "Small-volume extrusion apparatus for preparation of large, unilamellar vesicles," Biochim. Biophys.Acta. Biomembr. 1061, 297-303 (1991).
[CrossRef]

1989

P. J. Wyatt and C. Jackson, "Discrimination of Phytoplankton via light-scattering properties," Limnol. Oceanogr. 34, 96-112 (1989).
[CrossRef]

R. J. Olson, R. R. Zettler, and O. K. Anderson, "Discrimination of Eukaryotic Phytoplankton cell types from light scatter and autofluorescence properties measured by Flow Cytometry," Cytometry 10, 636-643 (1989).
[CrossRef] [PubMed]

1978

P. Latimer and P. Barber, "Scattering by ellipsoids of revolution a comparison of theoretical methods," J. Colloid Interface Sci. 63, 310-316 (1978).
[CrossRef]

1975

P. Latimer, "Light scattering by ellipsoids," J. Colloid Interface Sci. 53, 102-109 (1975).
[CrossRef]

1973

F. T. Gucker, J. Tuma, H. M. Lin, H. C. M., S. C. Ems, and T. R. Marshall, "Rapid measurement of light-scattering diagrams from single particles in an aerosol stream and dtermination of latex particle size," Aerosol Sci. 4, 389-404 (1973).
[CrossRef]

Anderson, O. K.

R. J. Olson, R. R. Zettler, and O. K. Anderson, "Discrimination of Eukaryotic Phytoplankton cell types from light scatter and autofluorescence properties measured by Flow Cytometry," Cytometry 10, 636-643 (1989).
[CrossRef] [PubMed]

Barber, P.

P. Latimer and P. Barber, "Scattering by ellipsoids of revolution a comparison of theoretical methods," J. Colloid Interface Sci. 63, 310-316 (1978).
[CrossRef]

Bernard, S.

A. Quirantes and S. Bernard, "Light scattering by marine algae: two-layer spherical and nonspherical models," J. Quant. Spectrosc. Radiat. Transfer 89,311-321 (2004).
[CrossRef]

Bricaud, A.

Chachisvilis, M.

D. Watson, N. Hagen, J. Diver, P. Marchand, and M. Chachisvilis, "Elastic light scattering from single cells: orientational dynamics in optical trap," Biophys. J. 87, 1298-1306 (2004).
[CrossRef] [PubMed]

Charlton, F.

H. Volten, J. F. de Haan, J. W. Hovenier, R. Schreurs, W. Vassen, A. G. Dekker, H. J. Hoogenboom, F. Charlton, R. Wouts, "Laboratory measurements of Angular Distributions of light scattered by Phytoplankton and Silt," Limnol. Oceanogr. 43, 1180-1197 (1998).
[CrossRef]

de Haan, J. F.

H. Volten, J. F. de Haan, J. W. Hovenier, R. Schreurs, W. Vassen, A. G. Dekker, H. J. Hoogenboom, F. Charlton, R. Wouts, "Laboratory measurements of Angular Distributions of light scattered by Phytoplankton and Silt," Limnol. Oceanogr. 43, 1180-1197 (1998).
[CrossRef]

de Marsac, N. T.

J. M. Garcia-Fernandez, N. T. de Marsac, and J. Diez, "Streamlined regulation and gene loss as adaptive mechanisms in prochlorococcus for optimized nitrogen utilization in Oligotrophic environments," Microbiol. Mol. Biol. Rev. 68, 630-638 (2004).
[CrossRef] [PubMed]

Dekker, A. G.

H. Volten, J. F. de Haan, J. W. Hovenier, R. Schreurs, W. Vassen, A. G. Dekker, H. J. Hoogenboom, F. Charlton, R. Wouts, "Laboratory measurements of Angular Distributions of light scattered by Phytoplankton and Silt," Limnol. Oceanogr. 43, 1180-1197 (1998).
[CrossRef]

Diez, J.

J. M. Garcia-Fernandez, N. T. de Marsac, and J. Diez, "Streamlined regulation and gene loss as adaptive mechanisms in prochlorococcus for optimized nitrogen utilization in Oligotrophic environments," Microbiol. Mol. Biol. Rev. 68, 630-638 (2004).
[CrossRef] [PubMed]

Diver, J.

D. Watson, N. Hagen, J. Diver, P. Marchand, and M. Chachisvilis, "Elastic light scattering from single cells: orientational dynamics in optical trap," Biophys. J. 87, 1298-1306 (2004).
[CrossRef] [PubMed]

Drezek, R.

Dunn, A.

Garcia-Fernandez, J. M.

J. M. Garcia-Fernandez, N. T. de Marsac, and J. Diez, "Streamlined regulation and gene loss as adaptive mechanisms in prochlorococcus for optimized nitrogen utilization in Oligotrophic environments," Microbiol. Mol. Biol. Rev. 68, 630-638 (2004).
[CrossRef] [PubMed]

Greenaway, R. S.

Z. Ulanowski, R. S. Greenaway, P. H. Kaye, and I. K. Ludlow, "Laser diffractometer for single-particle scattering measurements," Meas. Sci. Technol 13, 292-296 (2002).
[CrossRef]

Gucker, F. T.

F. T. Gucker, J. Tuma, H. M. Lin, H. C. M., S. C. Ems, and T. R. Marshall, "Rapid measurement of light-scattering diagrams from single particles in an aerosol stream and dtermination of latex particle size," Aerosol Sci. 4, 389-404 (1973).
[CrossRef]

Hagen, N.

D. Watson, N. Hagen, J. Diver, P. Marchand, and M. Chachisvilis, "Elastic light scattering from single cells: orientational dynamics in optical trap," Biophys. J. 87, 1298-1306 (2004).
[CrossRef] [PubMed]

Hoogenboom, H. J.

H. Volten, J. F. de Haan, J. W. Hovenier, R. Schreurs, W. Vassen, A. G. Dekker, H. J. Hoogenboom, F. Charlton, R. Wouts, "Laboratory measurements of Angular Distributions of light scattered by Phytoplankton and Silt," Limnol. Oceanogr. 43, 1180-1197 (1998).
[CrossRef]

Hovenier, J. W.

H. Volten, J. F. de Haan, J. W. Hovenier, R. Schreurs, W. Vassen, A. G. Dekker, H. J. Hoogenboom, F. Charlton, R. Wouts, "Laboratory measurements of Angular Distributions of light scattered by Phytoplankton and Silt," Limnol. Oceanogr. 43, 1180-1197 (1998).
[CrossRef]

Hu, L.-r.

R. C. MacDonald, R. I. MacDonald, B. P. M. Menco, K. Takeshita, N. K. Subbarao, and L.-r. Hu, "Small-volume extrusion apparatus for preparation of large, unilamellar vesicles," Biochim. Biophys.Acta. Biomembr. 1061, 297-303 (1991).
[CrossRef]

Jackson, C.

P. J. Wyatt and C. Jackson, "Discrimination of Phytoplankton via light-scattering properties," Limnol. Oceanogr. 34, 96-112 (1989).
[CrossRef]

Jacquet, S.

D. Marie, F. Partensky, S. Jacquet, and D. Vaulot, "Enumeration and cell cycle analysis of Natural Populations of Marine Picoplankton by Flow Cytometry using the nucleic acid stain SYBR Green I," Appl. Environ. Microbiol. 63, 186-193 (1997).
[PubMed]

Kasting, J. F.

J. F. Kasting and J. L. Siefert, "Life and the evolution of Earth's atmosphere," Science 296, 1066-1068 (2002)
[CrossRef] [PubMed]

Kaye, P. H.

Z. Ulanowski, R. S. Greenaway, P. H. Kaye, and I. K. Ludlow, "Laser diffractometer for single-particle scattering measurements," Meas. Sci. Technol 13, 292-296 (2002).
[CrossRef]

Latimer, P.

P. Latimer and P. Barber, "Scattering by ellipsoids of revolution a comparison of theoretical methods," J. Colloid Interface Sci. 63, 310-316 (1978).
[CrossRef]

P. Latimer, "Light scattering by ellipsoids," J. Colloid Interface Sci. 53, 102-109 (1975).
[CrossRef]

Lee, M. E.

M. E. Lee and M. R. Lewis, "A New Method for the Measurement of the Optical Volume Scattering Function in the Upper Ocean," J. Atmos Ocean. Tech. 20, 563-571 (2003).
[CrossRef]

Lewis, M. R.

M. E. Lee and M. R. Lewis, "A New Method for the Measurement of the Optical Volume Scattering Function in the Upper Ocean," J. Atmos Ocean. Tech. 20, 563-571 (2003).
[CrossRef]

Lin, H. M.

F. T. Gucker, J. Tuma, H. M. Lin, H. C. M., S. C. Ems, and T. R. Marshall, "Rapid measurement of light-scattering diagrams from single particles in an aerosol stream and dtermination of latex particle size," Aerosol Sci. 4, 389-404 (1973).
[CrossRef]

Ludlow, I. K.

Z. Ulanowski, R. S. Greenaway, P. H. Kaye, and I. K. Ludlow, "Laser diffractometer for single-particle scattering measurements," Meas. Sci. Technol 13, 292-296 (2002).
[CrossRef]

MacDonald, R. C.

R. C. MacDonald, R. I. MacDonald, B. P. M. Menco, K. Takeshita, N. K. Subbarao, and L.-r. Hu, "Small-volume extrusion apparatus for preparation of large, unilamellar vesicles," Biochim. Biophys.Acta. Biomembr. 1061, 297-303 (1991).
[CrossRef]

MacDonald, R. I.

R. C. MacDonald, R. I. MacDonald, B. P. M. Menco, K. Takeshita, N. K. Subbarao, and L.-r. Hu, "Small-volume extrusion apparatus for preparation of large, unilamellar vesicles," Biochim. Biophys.Acta. Biomembr. 1061, 297-303 (1991).
[CrossRef]

Marchand, P.

D. Watson, N. Hagen, J. Diver, P. Marchand, and M. Chachisvilis, "Elastic light scattering from single cells: orientational dynamics in optical trap," Biophys. J. 87, 1298-1306 (2004).
[CrossRef] [PubMed]

Marie, D.

D. Marie, F. Partensky, S. Jacquet, and D. Vaulot, "Enumeration and cell cycle analysis of Natural Populations of Marine Picoplankton by Flow Cytometry using the nucleic acid stain SYBR Green I," Appl. Environ. Microbiol. 63, 186-193 (1997).
[PubMed]

Menco, B. P. M.

R. C. MacDonald, R. I. MacDonald, B. P. M. Menco, K. Takeshita, N. K. Subbarao, and L.-r. Hu, "Small-volume extrusion apparatus for preparation of large, unilamellar vesicles," Biochim. Biophys.Acta. Biomembr. 1061, 297-303 (1991).
[CrossRef]

Morel, A.

Olson, R. J.

R. J. Olson, R. R. Zettler, and O. K. Anderson, "Discrimination of Eukaryotic Phytoplankton cell types from light scatter and autofluorescence properties measured by Flow Cytometry," Cytometry 10, 636-643 (1989).
[CrossRef] [PubMed]

Partensky, F.

D. Marie, F. Partensky, S. Jacquet, and D. Vaulot, "Enumeration and cell cycle analysis of Natural Populations of Marine Picoplankton by Flow Cytometry using the nucleic acid stain SYBR Green I," Appl. Environ. Microbiol. 63, 186-193 (1997).
[PubMed]

Quirantes, A.

A. Quirantes and S. Bernard, "Light scattering by marine algae: two-layer spherical and nonspherical models," J. Quant. Spectrosc. Radiat. Transfer 89,311-321 (2004).
[CrossRef]

Richards-Kortum, R.

Schreurs, R.

H. Volten, J. F. de Haan, J. W. Hovenier, R. Schreurs, W. Vassen, A. G. Dekker, H. J. Hoogenboom, F. Charlton, R. Wouts, "Laboratory measurements of Angular Distributions of light scattered by Phytoplankton and Silt," Limnol. Oceanogr. 43, 1180-1197 (1998).
[CrossRef]

Siefert, J. L.

J. F. Kasting and J. L. Siefert, "Life and the evolution of Earth's atmosphere," Science 296, 1066-1068 (2002)
[CrossRef] [PubMed]

Stramski, D.

Subbarao, N. K.

R. C. MacDonald, R. I. MacDonald, B. P. M. Menco, K. Takeshita, N. K. Subbarao, and L.-r. Hu, "Small-volume extrusion apparatus for preparation of large, unilamellar vesicles," Biochim. Biophys.Acta. Biomembr. 1061, 297-303 (1991).
[CrossRef]

Takeshita, K.

R. C. MacDonald, R. I. MacDonald, B. P. M. Menco, K. Takeshita, N. K. Subbarao, and L.-r. Hu, "Small-volume extrusion apparatus for preparation of large, unilamellar vesicles," Biochim. Biophys.Acta. Biomembr. 1061, 297-303 (1991).
[CrossRef]

Tuma, J.

F. T. Gucker, J. Tuma, H. M. Lin, H. C. M., S. C. Ems, and T. R. Marshall, "Rapid measurement of light-scattering diagrams from single particles in an aerosol stream and dtermination of latex particle size," Aerosol Sci. 4, 389-404 (1973).
[CrossRef]

Ulanowski, Z.

Z. Ulanowski, R. S. Greenaway, P. H. Kaye, and I. K. Ludlow, "Laser diffractometer for single-particle scattering measurements," Meas. Sci. Technol 13, 292-296 (2002).
[CrossRef]

Vassen, W.

H. Volten, J. F. de Haan, J. W. Hovenier, R. Schreurs, W. Vassen, A. G. Dekker, H. J. Hoogenboom, F. Charlton, R. Wouts, "Laboratory measurements of Angular Distributions of light scattered by Phytoplankton and Silt," Limnol. Oceanogr. 43, 1180-1197 (1998).
[CrossRef]

Vaulot, D.

D. Marie, F. Partensky, S. Jacquet, and D. Vaulot, "Enumeration and cell cycle analysis of Natural Populations of Marine Picoplankton by Flow Cytometry using the nucleic acid stain SYBR Green I," Appl. Environ. Microbiol. 63, 186-193 (1997).
[PubMed]

Volten, H.

H. Volten, J. F. de Haan, J. W. Hovenier, R. Schreurs, W. Vassen, A. G. Dekker, H. J. Hoogenboom, F. Charlton, R. Wouts, "Laboratory measurements of Angular Distributions of light scattered by Phytoplankton and Silt," Limnol. Oceanogr. 43, 1180-1197 (1998).
[CrossRef]

Watson, D.

D. Watson, N. Hagen, J. Diver, P. Marchand, and M. Chachisvilis, "Elastic light scattering from single cells: orientational dynamics in optical trap," Biophys. J. 87, 1298-1306 (2004).
[CrossRef] [PubMed]

Wouts, R.

H. Volten, J. F. de Haan, J. W. Hovenier, R. Schreurs, W. Vassen, A. G. Dekker, H. J. Hoogenboom, F. Charlton, R. Wouts, "Laboratory measurements of Angular Distributions of light scattered by Phytoplankton and Silt," Limnol. Oceanogr. 43, 1180-1197 (1998).
[CrossRef]

Wyatt, P. J.

P. J. Wyatt and C. Jackson, "Discrimination of Phytoplankton via light-scattering properties," Limnol. Oceanogr. 34, 96-112 (1989).
[CrossRef]

Zettler, R. R.

R. J. Olson, R. R. Zettler, and O. K. Anderson, "Discrimination of Eukaryotic Phytoplankton cell types from light scatter and autofluorescence properties measured by Flow Cytometry," Cytometry 10, 636-643 (1989).
[CrossRef] [PubMed]

Acta. Biomembr.

R. C. MacDonald, R. I. MacDonald, B. P. M. Menco, K. Takeshita, N. K. Subbarao, and L.-r. Hu, "Small-volume extrusion apparatus for preparation of large, unilamellar vesicles," Biochim. Biophys.Acta. Biomembr. 1061, 297-303 (1991).
[CrossRef]

Aerosol Sci.

F. T. Gucker, J. Tuma, H. M. Lin, H. C. M., S. C. Ems, and T. R. Marshall, "Rapid measurement of light-scattering diagrams from single particles in an aerosol stream and dtermination of latex particle size," Aerosol Sci. 4, 389-404 (1973).
[CrossRef]

Appl. Environ. Microbiol.

D. Marie, F. Partensky, S. Jacquet, and D. Vaulot, "Enumeration and cell cycle analysis of Natural Populations of Marine Picoplankton by Flow Cytometry using the nucleic acid stain SYBR Green I," Appl. Environ. Microbiol. 63, 186-193 (1997).
[PubMed]

Appl. Opt.

Biophys. J.

D. Watson, N. Hagen, J. Diver, P. Marchand, and M. Chachisvilis, "Elastic light scattering from single cells: orientational dynamics in optical trap," Biophys. J. 87, 1298-1306 (2004).
[CrossRef] [PubMed]

Cytometry

R. J. Olson, R. R. Zettler, and O. K. Anderson, "Discrimination of Eukaryotic Phytoplankton cell types from light scatter and autofluorescence properties measured by Flow Cytometry," Cytometry 10, 636-643 (1989).
[CrossRef] [PubMed]

J. Atmos Ocean. Tech.

M. E. Lee and M. R. Lewis, "A New Method for the Measurement of the Optical Volume Scattering Function in the Upper Ocean," J. Atmos Ocean. Tech. 20, 563-571 (2003).
[CrossRef]

J. Colloid Interface Sci.

P. Latimer, "Light scattering by ellipsoids," J. Colloid Interface Sci. 53, 102-109 (1975).
[CrossRef]

P. Latimer and P. Barber, "Scattering by ellipsoids of revolution a comparison of theoretical methods," J. Colloid Interface Sci. 63, 310-316 (1978).
[CrossRef]

J. Plankton Res.

Eyvind Aas, "Refractive index of phytoplankton derived from its metabolite composition," J. Plankton Res. 18,2223-2249 (1996).
[CrossRef]

J. Quant. Spectrosc. Radiat. Transfer

A. Quirantes and S. Bernard, "Light scattering by marine algae: two-layer spherical and nonspherical models," J. Quant. Spectrosc. Radiat. Transfer 89,311-321 (2004).
[CrossRef]

Limnol. Oceanogr.

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Figures (7)

Fig. 1.
Fig. 1.

The schematic diagram of RM – I, showing the modified laboratory coordinate system, the incident (I0), and the scattered (IS) beams. The orientation of the spheroid particle is determined by γ and ψ. Q is the scattering angle, and d is the angle of the spheroid orientation with respect to the bisectrix (BS).

Fig. 2.
Fig. 2.

A Schematic of the optical setup used to record ARLS from suspended particles.[22]

Fig. 3.
Fig. 3.

Scattering diagrams of suspension of vesicles with 30 nm nominal diameter in PBS1 solution. (a). Simulation based on extended GLMT of coated sphere. (b). Experimental curve

Fig. 4.
Fig. 4.

Scattering diagrams of polystyrene microspheres with diameter of 0.6 µm, 0.8 µm and 1.0 µm, respectively. (a) Simulation with GLMT on intermediate-sized sphere. (b) Experimental measurements on low-concentration suspensions

Fig. 5.
Fig. 5.

Scattering diagram of Synechococcus sp. strain CC9311. (a). Simulation with RM-I method on a homogenous spheroid with 0.8×0.8×2.0 µm outer dimensions (a=0.4 µm, v=2.5 as in the spheroid equation) and n=1.406 in seawater (n=1.339). (b). Experimental measurements on low-concentration suspensions of Synechococcus sp. strain CC9311.

Fig. 6.
Fig. 6.

Scattering diagram of Flavobacterium sp. strain ALC1. (a). Simulation with RM-I on a homogenous spheroid with 1.0×1.0×3.5 µm outer dimensions (a=0.5 µm, v=3.5 as in the spheroid equation), and n=1.406 in seawater (n=1.339). (b). Experimental measurements on low-concentration suspensions of Flavobacterium sp. strain ALC1.

Fig. 7.
Fig. 7.

Scattering diagram of Ostreococcus sp. strain CCE9901. (a). Simulation with extended RM-I on a coated spheroid with 1.0×1.0×2.0 µm outer dimensions (a=0.5 µm, v=2 as in the spheroid equation) and a concentric spherical core of radius 0.25 µm. (n=1.40 for the spherical core, n=1.37 for the shell) in seawater (n=1.339). (b). Experimental measurements on lowconcentration suspensions of Ostreococcus sp. strain CCE9901.

Tables (2)

Tables Icon

Table 1. Two typical cases of a coated spheroid model and the corresponding mathematical definitions used in the extended RM – I method. x 1 --- size parameter of the core. y 1 --- size parameter of the outer spheroid.

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Table 2. Scattering simulation models for marine picoplankton to be studied.

Equations (3)

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

δ = π arccos [ ( sin ψ cos γ ) 2 + ( cos ψ ) 2 · cos [ arctan ( tan ψ cos γ ) + π 2 Θ 2 ] ]
g = ( sin 2 δ + v 2 cos 2 δ ) 1 2
d Ω = sin ( ψ ) · d ψ · d γ = d cos ( ψ ) · d γ

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