Skip to main content
Log in

Size distribution of submicron particles by dynamic light scattering measurements: analyses considering normalization errors

  • Published:
European Biophysics Journal Aims and scope Submit manuscript

Abstract

Errors in the experimental baseline used to normalize dynamic light scattering data can seriously affect the size distribution resulting from the data analysis. A revised method, which incorporates the characteristics of this error into the size distribution algorithm CONTIN (Ruf 1989), is tested with experimental data of high statistical accuracy obtained from a sample of phospholipid vesicles. It is shown that the various commonly used ways of accumulating and normalizing dynamic light scattering data are associated with rather different normalization errors. As a consequence a variety of solutions differing in modality, as well as in width, are obtained on carrying out data analysis in the common way. It is demonstrated that a single monomodal solution is retrieved from all these data sets when the new method is applied, which in addition provides the corresponding baseline errors quantitatively. Furthermore, stable solutions are obtainable with data of lower statistical accuracy which results from measurements of shorter duration. The use of an additional parameter in data inversion reduces the occurrence of spurious peaks. This stabilizing effect is accompanied by larger uncertainties in the width of the size distribution. It is demonstrated that these uncertainties are reduced by nearly a factor of two on using the normalization error function instead of the ‘dust term’ option for the analysis of noisy data sets.

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

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Aragon SR, Pecora R (1976) Theory of dynamic light scattering from polydisperse systems. J Chem Phys 64:2395–2404

    Google Scholar 

  • Chu B (1983) Correlation function profile analysis in laser light scattering 1. General review on methods of data analysis. In: Earnshaw JC, Steer MW (eds) The application of laser light scattering to the study of biological motion, NATO Adv Sci Inst Series A, vol 59. Plenum Press, NewYork London, pp 53–76

    Google Scholar 

  • Degiorgio V, Lastovka JB (1971) Intensity-correlation spectroscopy. Phys Rev A 4:2033–2050

    Google Scholar 

  • Flamberg A, Pecora R (1984) Dynamic light scattering study of micelles in a high ionic strength solution. J Phys Chem 88:3026–3033

    Google Scholar 

  • Georgalis Y, Ruf H, Grell E (1987) Photon-correlation-spectroscopy of membrane model systems. In: Burgen ASV, Roberts GCK, Anner BM (eds) Topics in molecular pharmacology, vol 4. Elsevier, Amsterdam, pp 1–20

    Google Scholar 

  • Hallett FR, Watton J, Krygsman P (1991) Vesicle sizing number distributions by dynamic light scattering. Biophys J 59:357–363

    Google Scholar 

  • Hughes AJ, Jakeman E, Oliver CJ, Pike ER (1973) Photon-correlation spectroscopy: dependence of linewidth error on normalization, clip level, detector area, sample time and count rate. J Phys A: Math Nucl Gen 6:1327–1336

    Google Scholar 

  • Jakeman E (1974) Photon correlation. In: Cummins HZ, Pike ER (eds) Photon correlation and light beating spectroscopy, NATO Adv Study Inst Series B, vol 3. Plenum Press, New York London, pp 75–149

    Google Scholar 

  • Jakeman E, Pike ER, Swain S (1971) Statistical accuracy in the digital autocorrelation of photon counting fluctuations. J Phys A: Gen Phys 4:517–534

    Google Scholar 

  • Kojro Z (1990) Influence of statistical errors on size distributions obtained from dynamic light scattering data. Experimental limitations in size distribution determination. J Phys A: Math Gen 23:1363–1383

    Google Scholar 

  • Kojro Z, Lin SQ, Grell E, Ruf H (1989) Determination of internal volume and volume distribution of lipid vesicles from dynamic light scattering data. Biochim Biophys Acta 985:1–8

    Google Scholar 

  • Koppel DE (1972) Analysis of macromolecular polydispersity in intensity correlation spectroscopy: the method of cumulants. J Chem Phys 57:4814–4820

    Google Scholar 

  • Minims LT, Zampighi G, Nozaki Y, Tanford C, Reynolds JA (1981) Phospholipid vesicle formation and transmembrane protein incorporation using octyl glucoside. Biochemistry 20:833–840

    Google Scholar 

  • Morrison ID, Grabowski EF, Herb CA (1985) Improved techniques for particle size determination by quasi-elastic light scattering. Langmuir 1:496–501

    CAS  Google Scholar 

  • Oliver J (1981) Recent development in photon correlation and spectrum analysis techniques: 11. Information from photodetection spectroscopy. In: Chen SH, Chu B, Nossal R (eds) Scattering techniques applied to supramolecular and nonequilibrium systems, NATO Adv Sci Inst Series B, vol 73. Plenum Press, New York London, pp 121–160

    Google Scholar 

  • Pecora R, Aragon SR (1974) Theory of light scattering from hollow spheres. Chem Phys Lipids 13:1–10

    Google Scholar 

  • Peters R (1991) Noise on photon correlation functions and its effect on data reduction algorithms. In: Brown W (ed) Dynamic light scattering, the method and some application. Oxford University Press, Oxford (in press)

    CAS  PubMed  Google Scholar 

  • Phillips DL (1962) A technique for the numerical solution of certain integral equations of the first kind. J Ass Comput Mach 9:84–97

    Google Scholar 

  • Pike ER, Watson D, McNeil Watson F (1983) Analysis of polydisperse scattering data II. In: Dahneke BE (ed) Measurement of suspended particles by quasi-elastic light scattering. Wiley, New York, pp 107–128

    Google Scholar 

  • Provencher SW (1979) Inverse problems in polymer characterization: direct analysis of polydispersity with photon correlation spectroscopy. Makromol Chem 180:201–209

    Google Scholar 

  • Provencher SW (1982a) A constrained regularization method for inverting data represented by linear algebraic or integral equations. Comput Phys Commun 27:213–227

    Google Scholar 

  • Provencher SW (1982b) CONTIN: A general purpose constrained regularization programm for inverting noisy linear algebraic and integral equations. Comput Phys Commun 27:229–242

    Google Scholar 

  • Provencher SW (1982 c) CONTIN, Users Manual. EMBL technical report DA05. Heidelberg: European Molecular Biology Lab

  • Provencher SW, Hendrix J, De Maeyer L, Paulussen N (1978) Direct determination of molecular weight distributions of polystyrene in cyclohexane with photon correlation spectroscopy. J Chem Phys 69:4273–4276

    Google Scholar 

  • Pusey PN, Vaughan JM (1975) Light scattering and intensity fluctuation spectroscopy. In: Davies M (ed) Dielectric and related molecular processes, vol 2. The Chemical Society, London, pp 48–105

    Google Scholar 

  • Ruf H (1989) Effects of normalization errors on size distributions obtained from dynamic light scattering data. Biophys J 56:67–78

    Google Scholar 

  • Ruf H, Georgalis Y, Grell E (1989) Dynamic light scattering to determine size distributions of vesicles. In: Fleischer S, Fleischer B (eds) Methods in enzymology, biomembranes part S, vol 172. Academic Press, San Diego, pp 364–390

    Google Scholar 

  • Saleh BEA, Cardoso MF (1973) The effect of channel correlation on the accuracy of photon counting digital autocorrelators. J Phys A Math Nucl Gen 6:1897–1909

    Google Scholar 

  • Schätzel K (1987) Correlation techniques in dynamic light scattering. Appl Phys B 42:193–213

    Google Scholar 

  • Schätzel K (1990) Noise on photon correlation data: I. Autocorrelation functions. Quant Opt 2:287–305

    Google Scholar 

  • Schätzel K, Drewel M, Stimac S (1988) Photon correlation measurements at large lag times: improving statistical accuracy. J Mod Opt 35:711–718

    Google Scholar 

  • Siegert AJF (1943). MIT Rad Lab Rep No 465

  • Stelzer EHK, Ruf H, Grell E (1983) Analysis and resolution of polydisperse systems. In: Schulz-Dubois EO (ed) Photon correlation techniques, Springer Series in Optical Sciences, vol 6. Springer, Berlin Heidelberg New York, pp 329–334

    Google Scholar 

  • Stock RS, Ray WH (1985) Interpretation of photon correlation spectroscopy data: A comparison of analysis methods. J Polym Sci Polym Phys 23:1393–1447

    Google Scholar 

  • Vaidya RA, Mettille MJ, Hester RD (1987) A comparison of methods for determining macromolecular polydispersity from dynamic light scattering data. In: Provder T (ed) Particle size distribution, ACS Symposium Series, vol 332. American Chemical Society, Washington DC, pp 62–73

    Google Scholar 

  • Weiner BB, Tscharnuter WW (1987) Uses and abuses of photon correlation spectroscopy. In: Provder T (ed) Particle size distribution, ACS Symposium Series, vol 332. American Chemical Society, Washington DC, pp 48–61

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Dedicated to Manfred Eigen on the occasion of his 65th birthday

Offrint requests to: H. Ruf

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ruf, H., Grell, E. & Stelzer, E.H.K. Size distribution of submicron particles by dynamic light scattering measurements: analyses considering normalization errors. Eur Biophys J 21, 21–28 (1992). https://doi.org/10.1007/BF00195440

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00195440

Key words

Navigation