DYNAMIC LIGHT
SCATTERING (DLS)
Presented by MUDHA TARIQ
Reg no MPH243010
OUTLINE
• What is DLS?
• Principle of DLS
• How DLS work ?
• Applications
• Advantages
• Limitations
Introduction
• Dynamic Light Scattering (DLS) is a technique used to
measure the size distribution of small particles in
suspension or polymers in solution by analyzing the
fluctuations in the intensity of scattered light.
• It is commonly used in the characterization of
nanoparticles, proteins, and other macromolecules.
Principle of DLS
• The principle of Dynamic Light Scattering (DLS) is based on analyzing the
fluctuations in the intensity of light scattered by particles in motion (due to
Brownian motion) within a sample. Here's a breakdown of how DLS works
in principle:
• Laser Illumination and Scattering
• Brownian Motion of Particles
• Autocorrelation Function
• Fluctuations in Scattered Light
• Stokes-Einstein Equation
• Stokes-Einstein Equation
• Hydrodynamic Diameter
Continue ….
1. Laser Illumination and Scattering
• A monochromatic laser light is directed at the sample, typically a suspension of particles or molecules in a liquid.
• When the laser light hits the particles, the particles scatter the light in different directions. The amount of light scattered
depends on the size of the particles.
2. Brownian Motion of Particles
• The particles in the sample are in constant motion due to thermal energy, a phenomenon known as Brownian motion.
• Smaller particles move faster than larger particles due to their lower mass, whereas larger particles move more slowly.
3. Fluctuations in Scattered Light
• The random movement (Brownian motion) of particles causes fluctuations in the intensity of scattered light over time.
• The detector collects these intensity fluctuations, which are related to how quickly the particles move.
4. Autocorrelation Function
• The detected intensity fluctuations are analyzed using an autocorrelation function, which compares the intensity at
one point in time with the intensity at a later point in time.
• The autocorrelation function provides information on how the intensity of the scattered light changes over time due to
the movement of particles.
Continue …
Stokes-Einstein Equation
• From the diffusion coefficient DDD, the particle size can be determined using the
Stokes-Einstein equation:
D=6πη/rkBT
Where:
• D is the diffusion coefficient of the particles.
• kBis the Boltzmann constant.
• T is the temperature in Kelvin.
• η is the viscosity of the solvent.
• r is the hydrodynamic radius of the particles.
• The particle size (or hydrodynamic radius) can be calculated by rearranging this
Continue…..
• Hydrodynamic Diameter
• The hydrodynamic diameter is the effective size of the particle in solution, which is
measured in DLS. It accounts for the particle and its surrounding solvent shell (due to
interactions between the particle and solvent molecules).
• The result from DLS is usually a distribution of sizes (since the sample might contain
particles of different sizes) that can be interpreted as a histogram of particle sizes.
• Key Considerations:
• Wavelength of the laser affects resolution (shorter wavelengths are better for smaller
particles).
• Detection angle influences the sensitivity to different sizes (small angles are better for
larger particles).
How DLS works:
i. Laser and Scattering: A laser light is directed at the sample, and the particles in the
sample scatter the light in different directions.
ii. Brownian Motion: The particles undergo Brownian motion (random movement due to
thermal energy). This motion causes fluctuations in the intensity of the scattered light.
iii. Measurement of Fluctuations: A detector captures these intensity fluctuations over
time. The rate at which these fluctuations occur depends on the size of the particles:
Smaller particles: Move faster, causing more rapid fluctuations in the scattered light.
Larger particles: Move more slowly, leading to slower fluctuations.
iv. Calculation of Size: The technique uses the autocorrelation function to analyze the
fluctuations in light scattering. From this, the diffusion coefficient of the particles can
be determined, which is then related to the particle size using the Stokes-Einstein
equation.
AUTOCORRELATION
• Understanding Autocorrelation
Autocorrelation Function:
The autocorrelation function g(t)g(t) compares light intensity at one point in time
with that at a later time
Decay of autocorrelation: Faster decay implies smaller particles (due to quicker
motion), while slower decay corresponds to larger particles.
Time Scale: The characteristic time of decay is related to particle diffusion and thus
their size.
DLS Instrumentation
Components:
Laser source for light scattering.
Detector to measure the scattered light
intensity.
Autocorrelation function analysis to
determine particle size.
Applications of Dynamic Light Scattering
(DLS)
Nanoparticle Characterization
•Measures particle size, size distribution, and stability in suspensions.
•Used in nanomedicine, electronics, and materials science.
Pharmaceuticals & Drug Delivery
•Characterizes drug carriers (e.g., liposomes, nanoparticles).
•Studies protein aggregation and formulation stability.
Colloidal Systems & Suspensions
•Monitors colloidal stability (e.g., emulsions, dispersions).
•Used in food, cosmetics, and industrial formulations.
Biotechnology & Protein Analysis
•Analyzes protein aggregation, folding, and molecular interactions.
•Studies enzyme activity and drug-protein interactions.
Applications of DLS
Polymer & Macromolecule Studies
•Measures polymer size distribution and behavior in solution.
•Characterizes polymer blends and nanocomposites
Environmental Monitoring
•Analyzes nanoparticles in environmental samples (e.g., water, air).
•Monitors particulate matter for air quality control.
Aerosols & Particulate Matter
•Characterizes particle size in aerosols for inhalation therapy.
•Measures particulate matter for air pollution studies.
Industrial Quality Control
•Monitors particle size during manufacturing processes.
•Ensures consistent quality in products like paints and coatings.
Advantages of DLS
•Size Range Limitations
•Less accurate for particles larger than a few micrometers or very small particles below 1 nm.
•Multiple Scattering Effects
•High particle concentrations can lead to multiple scattering, distorting size measurements.
•Sensitivity to Sample Heterogeneity
•Difficult to accurately measure samples with a broad or complex size distribution.
•Dilution Requirement
•Requires careful sample dilution to avoid multiple scattering and ensure reliable results.
•Assumption of Spherical Particles
•Assumes spherical particles for calculations, which can lead to inaccuracies for irregularly
shaped particles.
•Sensitivity to Temperature and Viscosity
•Measurements can be affected by fluctuations in temperature or solvent viscosity.
Limitations of DLS
•Size Range Limitations
•Less accurate for particles larger than a few micrometers or very small particles below 1 nm.
•Multiple Scattering Effects
•High particle concentrations can lead to multiple scattering, distorting size measurements.
•Sensitivity to Sample Heterogeneity
•Difficult to accurately measure samples with a broad or complex size distribution.
•Dilution Requirement
•Requires careful sample dilution to avoid multiple scattering and ensure reliable results.
•Assumption of Spherical Particles
•Assumes spherical particles for calculations, which can lead to inaccuracies for irregularly shaped particles.
•Sensitivity to Temperature and Viscosity
•Measurements can be affected by fluctuations in temperature or solvent viscosity.
Conclusion
Summary:
DLS is a powerful, non-destructive method for determining particle size and
distribution across multiple industries, from pharmaceuticals to nanotechnology.
•Key Points:
• Fast, sensitive, and versatile.
• Ideal for nanoparticle and protein characterization.
• Limited by sample homogeneity and size range.
References
• Malvern Panalytical [Link]
• https://
[Link]/publication/331022012_Dynamic_lig
ht_scattering_DLS_technique_principle_theoretical_consider
ations_and_applications
• Falke S, Betzel C. Dynamic Light Scattering (DLS):
Principles, Perspectives, Applications to Biological Samples.
Radiation in Bioanalysis. 2019 Jul 29;8:173–93. doi:
10.1007/978-3-030-28247-9_6. PMCID: PMC7120185.