## Particle aggregation monitoring by speckle size measurement; application to blood platelets aggregation

Optics Express, Vol. 12, Issue 19, pp. 4596-4601 (2004)

http://dx.doi.org/10.1364/OPEX.12.004596

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### Abstract

We report on a new method based on speckle size analysis and devoted to particle aggregation measurements. The experimental measurements give the speckle size variation during a salt aggregation process of polystyrene microspheres. The measurements are taken at a fixed monomer concentration, varying the salt concentration. Moreover, we applied this technique to follow blood platelet aggregation, usually monitored with a visible light transmittance photometer (aggregometer). Aggregation process was induced by ADP (adenosine diphosphate) addition, then we measured the speckle size variation versus time at two different ADP concentrations.

© 2004 Optical Society of America

## 1. Introduction

1. J.P. Wilcoxon, J.E. Martin, and D.W. Shaefer, “Aggregation in colloidal gold,” Phys. Rev. A **39**, 2675–2688 (1989). [CrossRef] [PubMed]

2. M. Carpinetti, F. Ferri, M. Giglio, E. Paganini, and U. Perini, “Salt-induced fast aggregation of polystyrene latex,” Phys. Rev. A **42**, 7347–7354 (1990). [CrossRef]

3. Y. Georgalis, E.B. Starikov, B. Hollenbach, R. Lurz, Eberhard Scherzinger, W. Sger, H. Lehrach, and E. Wanker, “Huntingtin aggregation monitored by dynamic light scattering,” Proc. Natl. Acad. Sci. **95**, 6118–6121 (1998). [CrossRef] [PubMed]

4. D. Majolino, F. Mallamace, P. Migliardo, N. Micali, and C. Vasi, “Elastic and quasielastic light-scattering studies of the aggregation phenomena in water solutions of polystyrene particles,” Phys. Rev. A **40**, 4665–4674 (1989). [CrossRef] [PubMed]

5. D.A. Boas and A. G. Yodh, “Spatially varying dynamical properties of turbid media probed with diffusing temporal light correlation,” J. Opt. Soc. Am. A **14**, 192–215 (1997). [CrossRef]

7. P. Lehmann, “Surface-roughness measurement based on the intensity correlation function of scattered light under speckle-pattern illumination,” Appl. Opt. **38**, 1144–1152 (1999). [CrossRef]

8. G. Da Costa and J. Ferrari, “Anisotropic speckle patterns in the light scattered by rough cylindrical surfaces,” Appl. Opt. **36**, 5231–5237 (1997). [CrossRef] [PubMed]

9. R. Berlasso, F. Perez Quintian, M.A. Rebollo, C.A. Raffo, and N.G. Gaggioli, “Study of speckle size of light scattered from cylindrical rough surfaces,” Appl. Opt. **39**, 5811–5819 (2000). [CrossRef]

10. A. Sadhwani, K.T. Schomaker, G.J. Tearney, and N.S. Nishioka, “Determination of Teflon thickness with laser speckle. I. Potential for burn depth diagnosis,” Appl. Opt. **35**, 5727–5735 (1996). [CrossRef] [PubMed]

2. M. Carpinetti, F. Ferri, M. Giglio, E. Paganini, and U. Perini, “Salt-induced fast aggregation of polystyrene latex,” Phys. Rev. A **42**, 7347–7354 (1990). [CrossRef]

12. P. Latimer, “Blood platelet aggrgometer: predicted effects of aggregation, photometer geometry, and multiple scattering,” Appl. Opt. **22**, 1136–1143 (1983). [CrossRef] [PubMed]

## 2. Methodology and speckle size measurement

13. Y. Piederriere, J. Cariou, Y. Guern, B. Le Jeune, G. Le Brun, and J. Lotrian, “Scattering through fluids: speckle size measurement and Monte Carlo simulations close to and into the multiple scattering,” Opt. Express **12**, 176–188 (2004), http://www.opticsexpress.org/abstract.cfm?URI=OPEX-12-1-176 [CrossRef] [PubMed]

*μ*, on the speckle size. In the previous investigations about the speckle produced in transmission by scattering fluid media, we followed its evolution from size measurement versus the optical thickness,

_{s}*ls*=

*Lμ*(

_{s}*L*is the thickness of the slab), and versus the polystyrene-microsphere size in the media. The different optical thicknesses were obtained by varying the diffuser number,

*N*, since

*μ*is proportional to

_{s}*N*[14] for a given particle size. At weak scattering the speckle size decreases linearly with scattering increase. Moreover, for the biggest particles and a fixed optical thickness, the speckle size increases concomitantly with the particle diameter or diffuser size,

*d*. This variation was measured for diameters within 0.53 and 6.36 μm.

*N*and

*d*vary, and then both produce an increase of the speckle size: the sticking between particle induces an increase of

*d*and a decrease of

*N*.

*x*,

*y*). This function corresponds to the normalized autocorrelation function of the intensity; it has a zero base and its width provides a reasonable measurement of the “average width” of a speckle [6]. If

*I*(

*x*

_{1},

*y*

_{1}) and

*I*(

*x*

_{2},

*x*

_{2})are the intensities of two points in the observation plane (

*x*,

*y*), the intensity autocorrelation function is defined by Eq. (1) [6]:

*x*=

*x*

_{1}-

*x*

_{2}and Δ

*y*=

*y*

_{1}-

*y*

_{2}. ⟨ ⟩ corresponds to a spatial average. If

*x*

_{2}= 0,

*y*

_{2}= 0,

*x*

_{1}=

*x*and

*y*

_{1}=

*y*, we can write:

*FT*

^{-1}) of the Power Spectral Density (PSD) of the intensity:

*c*(

_{I}*x*,

*y*) calculated from the intensity distribution measured of the speckle is:

*c*(

_{I}*x*, 0) and

*c*(0,

_{I}*y*) are the horizontal and the vertical profile of

*c*(

_{I}*x*,

*y*), respectively. Let us term

*dx*the width of

*c*(

_{I}*x*,0) so that

*c*(

_{I}*dx*/2,0) = 0.5 and

*dy*the width of

*c*(0,

_{I}*y*) such as

*c*(0,

_{I}*dy*/2) = 0.5.

*I*/

_{0}*e*with

^{2}*I*being the maximum laser intensity, at the 632.8 nm wavelength with a coherence length of about 20 cm. The light intensity is monitored by a half wave plate

_{0}*L*associated to a polarizer P1. The length

*L*of the fluid sample is 1 cm.

^{-3}to 10

^{-4}s. However, in order to keep a correct signal-to-noise ratio, in our experiments image acquisition took 0.001 s.

15. Terri L. Alexander, James E. Harvey, and Arthur R. Weeks, “Average speckle size as a function of intensity threshold level: comparison of experimental measurements with theory,” Appl. Opt. **33**, 8240–8250 (1994). [CrossRef] [PubMed]

*D*(

*D*= 36 cm) between the sample and the CCD camera.

*θ*with the optical axis (Fig. 1). In our previous study [13

13. Y. Piederriere, J. Cariou, Y. Guern, B. Le Jeune, G. Le Brun, and J. Lotrian, “Scattering through fluids: speckle size measurement and Monte Carlo simulations close to and into the multiple scattering,” Opt. Express **12**, 176–188 (2004), http://www.opticsexpress.org/abstract.cfm?URI=OPEX-12-1-176 [CrossRef] [PubMed]

*dx*, when

*θ*< 10° dy variation versus

*θ*is small. In our experiments we have

*θ*= 5°.

## 3. Polystyrene microsphere aggregation

*N*, is known. The undiluted solution contains

_{0}*N*

_{0}=3.147 * 10

^{17}m

^{-3}. By dilution in deionized water, we adjusted the parameter

*N*and so the scattering coefficient.

*μ*determined by Beer-Lambert Law application:

^{s}*μ*= 6.23

_{s}*c*, where

*c*is the concentration of the initial solution. Furthermore, we employed deionized water to avoid the possible effect of particle aggregation due to electrostatic interaction. The refractive indices were 1.59 and 1.33 for the spheres and medium, respectively.

*μ*= 4 cm

_{s}^{-1}, and the initial speckle size measured about 46 μm. For our experimental configuration, the speckle size cannot exceed about 220 μm.

*dy*220 μm when

*μ*0 cm

_{s}^{-1}and this speckle size limit depends on the intensity distribution of the laser beam size [13

13. Y. Piederriere, J. Cariou, Y. Guern, B. Le Jeune, G. Le Brun, and J. Lotrian, “Scattering through fluids: speckle size measurement and Monte Carlo simulations close to and into the multiple scattering,” Opt. Express **12**, 176–188 (2004), http://www.opticsexpress.org/abstract.cfm?URI=OPEX-12-1-176 [CrossRef] [PubMed]

_{2}. Figure 2 illustrates

*dy*evolution versus the time,

*t*, for several salt concentrations.

*dy*increases a lot after the salt addition. Speckle size varies from 46 to 126 μm, which corresponds to a fast aggregation. After this first variation,

*dy*evolution is insignificant. At the weakest salt addition (3 mM), there is no strong variation. The small speckle size evolution corresponds to a slow aggregation. For the intermediary salt addition (45 mM), we can observed a strong variation of

*dy*next the salt addition and afterward we observe a weak variation: the aggregation process is fast further to the salt addition, then slows down.

## 4. Blood platelets aggregation

^{-1}and the speckle size before aggregation

*dy*= 100 μm. Large diffusers induce a large speckle size [13

**12**, 176–188 (2004), http://www.opticsexpress.org/abstract.cfm?URI=OPEX-12-1-176 [CrossRef] [PubMed]

*dy*is higher even though

*μs*is higher (platelet size is about 4 μm and polystyrene microsphere size is 0.535 μm).

*dy*from 100 to 135 μm). Afterwards, one should note the slow increase of the speckle size concomitant with the slowing down of the aggregation process.

*t*= 1.5 min.

## 5. Conclusion

12. P. Latimer, “Blood platelet aggrgometer: predicted effects of aggregation, photometer geometry, and multiple scattering,” Appl. Opt. **22**, 1136–1143 (1983). [CrossRef] [PubMed]

## References and links

1. | J.P. Wilcoxon, J.E. Martin, and D.W. Shaefer, “Aggregation in colloidal gold,” Phys. Rev. A |

2. | M. Carpinetti, F. Ferri, M. Giglio, E. Paganini, and U. Perini, “Salt-induced fast aggregation of polystyrene latex,” Phys. Rev. A |

3. | Y. Georgalis, E.B. Starikov, B. Hollenbach, R. Lurz, Eberhard Scherzinger, W. Sger, H. Lehrach, and E. Wanker, “Huntingtin aggregation monitored by dynamic light scattering,” Proc. Natl. Acad. Sci. |

4. | D. Majolino, F. Mallamace, P. Migliardo, N. Micali, and C. Vasi, “Elastic and quasielastic light-scattering studies of the aggregation phenomena in water solutions of polystyrene particles,” Phys. Rev. A |

5. | D.A. Boas and A. G. Yodh, “Spatially varying dynamical properties of turbid media probed with diffusing temporal light correlation,” J. Opt. Soc. Am. A |

6. | J.W. Goodman, “Statistical Properties of Laser Speckle Patterns,” in |

7. | P. Lehmann, “Surface-roughness measurement based on the intensity correlation function of scattered light under speckle-pattern illumination,” Appl. Opt. |

8. | G. Da Costa and J. Ferrari, “Anisotropic speckle patterns in the light scattered by rough cylindrical surfaces,” Appl. Opt. |

9. | R. Berlasso, F. Perez Quintian, M.A. Rebollo, C.A. Raffo, and N.G. Gaggioli, “Study of speckle size of light scattered from cylindrical rough surfaces,” Appl. Opt. |

10. | A. Sadhwani, K.T. Schomaker, G.J. Tearney, and N.S. Nishioka, “Determination of Teflon thickness with laser speckle. I. Potential for burn depth diagnosis,” Appl. Opt. |

11. | Y. Ozaki, “Measurement of platelet aggregation and attempts for standardization,” Sysmex J. Int. |

12. | P. Latimer, “Blood platelet aggrgometer: predicted effects of aggregation, photometer geometry, and multiple scattering,” Appl. Opt. |

13. | Y. Piederriere, J. Cariou, Y. Guern, B. Le Jeune, G. Le Brun, and J. Lotrian, “Scattering through fluids: speckle size measurement and Monte Carlo simulations close to and into the multiple scattering,” Opt. Express |

14. | H.C. Van de Hulst, |

15. | Terri L. Alexander, James E. Harvey, and Arthur R. Weeks, “Average speckle size as a function of intensity threshold level: comparison of experimental measurements with theory,” Appl. Opt. |

**OCIS Codes**

(030.6140) Coherence and statistical optics : Speckle

(120.3890) Instrumentation, measurement, and metrology : Medical optics instrumentation

(170.1470) Medical optics and biotechnology : Blood or tissue constituent monitoring

(170.4580) Medical optics and biotechnology : Optical diagnostics for medicine

(290.5850) Scattering : Scattering, particles

**ToC Category:**

Research Papers

**History**

Original Manuscript: July 26, 2004

Revised Manuscript: August 23, 2004

Published: September 20, 2004

**Citation**

Yann Piederrière, J. Le Meur, J. Cariou, J. Abgrall, and M. Blouch, "Particle aggregation monitoring by speckle size measurement; application to blood platelets aggregation," Opt. Express **12**, 4596-4601 (2004)

http://www.opticsinfobase.org/oe/abstract.cfm?URI=oe-12-19-4596

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### References

- J.P. Wilcoxon, J.E. Martin, D.W. Shaefer, �??Aggregation in colloidal gold,�?? Phys. Rev. A 39, 2675-2688 (1989). [CrossRef] [PubMed]
- M. Carpinetti, F. Ferri, M. Giglio, E. Paganini, U. Perini, �??Salt-induced fast aggregation of polystyrene latex,�?? Phys. Rev. A 42, 7347-7354 (1990). [CrossRef]
- Y. Georgalis, E.B. Starikov, B. Hollenbach, R. Lurz, Eberhard Scherzinger, W. Sger, H. Lehrach, E. Wanker, �??Huntingtin aggregation monitored by dynamic light scattering,�?? Proc. Natl. Acad. Sci. 95, 6118-6121 (1998). [CrossRef] [PubMed]
- D. Majolino, F. Mallamace, P. Migliardo, N. Micali, C. Vasi, �??Elastic and quasielastic light-scattering studies of the aggregation phenomena in water solutions of polystyrene particles,�?? Phys. Rev. A 40, 4665-4674 (1989). [CrossRef] [PubMed]
- D.A. Boas and A. G. Yodh, �??Spatially varying dynamical properties of turbid media probed with diffusing temporal light correlation,�?? J. Opt. Soc. Am. A 14, 192-215 (1997). [CrossRef]
- J.W. Goodman, �??Statistical Properties of Laser Speckle Patterns,�?? in Laser speckle and related phenomena, Vol. 9 in series Topics in Applied Physics, J.C. Dainty, Ed., (Springer-Verlag, Berlin, Heidelberg New York Tokyo, 1984).
- P. Lehmann, �??Surface-roughness measurement based on the intensity correlation function of scattered light under speckle-pattern illumination,�?? Appl. Opt. 38, 1144-1152 (1999). [CrossRef]
- G. Da Costa and J. Ferrari, �??Anisotropic speckle patterns in the light scattered by rough cylindrical surfaces,�?? Appl. Opt. 36, 5231-5237 (1997). [CrossRef] [PubMed]
- R. Berlasso, F. Perez Quintian, M.A. Rebollo, C.A. Raffo and N.G. Gaggioli, �??Study of speckle size of light scattered from cylindrical rough surfaces,�?? Appl. Opt. 39, 5811-5819 (2000). [CrossRef]
- A. Sadhwani, K.T. Schomaker, G.J. Tearney and N.S. Nishioka, �??Determination of Teflon thickness with laser speckle. I. Potential for burn depth diagnosis,�?? Appl. Opt. 35, 5727-5735 (1996). [CrossRef] [PubMed]
- Y. Ozaki, �??Measurement of platelet aggregation and attempts for standardization,�?? Sysmex J. Int. 8, 15-22 (1998).
- P. Latimer, �??Blood platelet aggrgometer: predicted effects of aggregation, photometer geometry, and multiple scattering,�?? Appl. Opt. 22, 1136-1143 (1983). [CrossRef] [PubMed]
- Y. Piederriere, J. Cariou, Y. Guern, B. Le Jeune, G. Le Brun, J. Lotrian, �??Scattering through fluids: speckle size measurement and Monte Carlo simulations close to and into the multiple scattering,�?? Opt. Express 12, 176-188 (2004), <a href="http://www.opticsexpress.org/abstract.cfm?URI=OPEX-12-1-176">http://www.opticsexpress.org/abstract.cfm?URI=OPEX-12-1-176</a> [CrossRef] [PubMed]
- H.C. Van de Hulst, Light scattering by small particles (New York, Dover, 1981).
- Terri L. Alexander, James E. Harvey, and Arthur R. Weeks, �??Average speckle size as a function of intensity threshold level: comparison of experimental measurements with theory,�?? Appl. Opt. 33, 8240-8250 (1994). [CrossRef] [PubMed]

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