Wyatt Technology

Widely used in biochemistry, biotechnology and pharmaceutical development, Dynamic Light Scattering (DLS) is popular wherever the size distributions of macromolecules and nanoparticles need to be measured quickly and easily.

Dynamic Light Scattering

Size distributions without fractionation

Dynamic Light Scattering (DLS) measures the translational diffusion coefficients Dt of nanoparticles and colloids in solution by quantifying dynamic fluctuations in scattered light. Sizes and size distributions, in turn, are calculated from the diffusion coefficients in terms of hydrodynamic radius rh or hydrodynamic diameter dh.

DLS is suitable for ensemble measurements ranging from rh values of 0.2 nm up to 5,000 nm. Wyatt offers high-throughput, automated DLS as well as conventional, cuvette-based formats.

Applications of DLS

Here are just a few of the diverse applications supported by DLS:

  • Estimate populations of aggregates large and small, for small molecules, proteins or liposomes
  • Check quality of biomolecules prior to running costly analyses such as SPR or SANS
  • Analyze thermal stability and chemical denaturation, differentiating pure unfolding from aggregation
  • Determine sizes of metallic nanoparticles or quantum dots
  • Quantify self-assembly processes of polypeptides or DNA
  • Assess colloidal stability as a precursor to aggregation and precipitation
  • Measure viscosity of concentrated biotherapeutics with just a few microliters of solution

DLS analysis of monomer

DLS analysis showing a protein monomer mixed with 50 nm particulates. The size resolution of unfractionated solutions is a factor of 3-5x in radius.

Dispersity

DLS determines size distributions without fractionation, providing polydispersity estimates as well as hydrodynamic radii.

Dynamic Light Scattering is also known as Quasi-Elastic Light Scattering (QELS) and Photon Correlation Spectroscopy (PCS).

Automated DLS

Low-volume, automated measurements performed directly in place in standard 96-, 384- or 1536-well microwell plates reveal new horizons for dynamic light scattering. Some of the high-throughput screening tasks accomplished with the DynaPro Plate Reader III include:

  • Quantifying type and degree of aggregation in hundreds of buffer conditions, important in formulation and crystallization studies
  • Pre-formulation of biotherapeutic candidates to evaluate developability
  • Assessment of buffer excipients and pH for colloidal stability using the diffusion interaction parameter kD
  • Assessment of multiple formulations for thermal (conformational) stability using the melting temperature Tm
  • Screening compound libraries for inhibition or enhancement of protein-protein interactions
  • Integration of plate-based testing protocols with other kinds of plate readers

In addition, microwell-plate-based DLS is an attractive option for multi-user labs: with no costly quartz cuvettes to wash, each user can have an inexpensive, personal, disposable plate sufficient for hundreds of samples and no concern for cross-contamination with other experiments.

Melting/aggregation curve data and analyses for four proteins measured simultaneously, in triplicate, on the DynaPro Plate Reader III.

Color-coded Results

Color-coded results of measurements of three proteins, four pH conditions and 6 concentrations, with five replicates per combination, in a 384 well plate. Total measurement time was < 1.5 hours.

Selected references

  Afonin, K. A.; Kasprzak, W.; Bindewald, E.; Puppala, P. S.; Diehl, A. R.; Hall, K. T.; Kim, T. J.; Zimmermann, M. T.; Jernigan, R. L.; Jaeger, L.; Shapiro, B. A. Computational and experimental characterization of RNA cubic nanoscaffolds. Methods  2014, 67, 256-265.

  Akiyama, S. Quality control of protein standards for molecular mass determinations by small-angle X-ray scattering. J. Appl. Crystallogr.  2010, 43, 237-243.

  An, W.; Zhang, H.; Sun, L.; Hao, A.; Hao, J.; Xin, F. Reversible vesicles based on one and two head supramolecular cyclodextrin amphiphile induced by methanol. Carbohyd. Res.  2010, 345, 914-921.

  Ferré-D'Amare, A. R.; Burley, S. K. Dynamic light scattering in evaluating crystallizability of macromolecules. Method. Enzymol.  1997, 276, 157-166.

  Mohr, J.; Chuan, Y. P.; Wu, Y.; Lua, L. H. L.; Middelberg, A. P. J. Virus-like particle formulation optimization by miniaturized high-throughput screening. Methods  2013, 60, 248-256.

 BIBLIOGRAPHIC SEARCH

Application notes

Instrumentation for DLS

  • Cuvette-based DLS Instrument

    DynaPro NanoStar, molar mass determination, molar mass measurement, molar mass analysis, molecular weight measurement, molar mass 
              characterization

    DynaPro® NanoStar® - With sample volumes as small as 1.25 µL and temperature control spanning -15°C to +150°C, the NanoStar goes above and beyond traditional cuvette-based DLS instruments. It offers an optimized static light scattering detector in parallel to the DLS detection system in order to determine true molar mass.

  • Automated DLS Instrument

    DynaPro® Plate Reader II - The only commercially available instrument that measures DLS directly in situ  in standard 96-, 384- or 1536-microwell plates, the non-perturbing DynaPro Plate Reader II offers integration with robot liquid handlers and multi-technique plate-based assay protocols. Temperature-controlled from 4°C to 85°C. An integrated camera views each well for troubleshooting and diagnostics.
    DynaPro Plate Reader II, DLS, sec analysis, quasi elastic light scattering, DynaPro NanoStar, QELS
    Möbiuζ The Möbiuζ is not only the most sensitive zeta potential detector available, it is also a top-of-the-line dynamic light scattering detector. The Möbiuζ provides connectivity to an autosampler for hands off, multi-sample operation.

    The Möbiuζ is also the only mobility detector that measures dynamic light scattering (DLS) simultaneously with zeta potential, in the same scattering volume, to monitor potential sample degradation.

    mobius-atlas, MP-PALS, zeta potential, zeta potential instrument, zetasizer, electrophoretic mobility, 
              isoelectric point, dn/dc
  • On-Line DLS Detector

    WyattQELS, accessory, DLS, sec analysis, quasi elastic light scattering, DynaPro Plate Reader II

    WyattQELS™ - A dynamic light scattering (DLS) module which integrates into a DAWN or miniDAWN MALS detector to provide simultaneous DLS measurements in the same scattering volume as MALS measurements.

  • Accessories

    microCUVETTE

    microCUVETTE - Allows static and/or dynamic light scattering measurements to be made in a quartz cuvette in a DAWN, requiring only 10 µL.

    NanoFilter Kit - Makes it easy to filter, recover and refilter precious sample. This is especially useful when your total sample volumes are below 300 µL.

   

For essential macromolecular and nanoparticle characterization—The Solution is Light™

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Wyatt Technology is the recognized leader in light scattering instrumentation and software for determining the absolute molar mass, size, charge and interactions of macromolecules and nanoparticles in solution.

Wyatt's line of multi-angle static light scattering products couple to size exclusion chromatography (SEC-MALS), field-flow fractionation (FFF-MALS), and stop-flow composition-gradient systems (CG-MALS). Our dynamic light scattering (DLS) products operate in traditional cuvette as well as on-line and automated, high-throughput modes. We also offer unique instruments for electrophoretic light scattering (MP-PALS), differential refractometry, and differential viscosity.



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