Dynamic light
scattering for size
determination
By,
Aniruddha Newaskar
Omprakash Sharma
Mohit Gupta
Neeraj Maurya
Outline
What is DLS
Applications of DLS
Brownian motion
Stoke’s Einstein equation : Relating particle size to particle
motion
How DLS works
The correlation function
How correlator works
Comparative Techniques
Limitations of DLS
Summary
What is Dynamic light Scattering
• Particle size can be determined by measuring the random
change in intensity of light scattered from suspension.
• It measure and interpolate the light scattering up to
microsecond
• So it measure real time intensity, thus measuring the dynamic
properties
Size distribution
Hydrodynamic radius
Diffusion coefficient
pm nm µm mm
1x10-1 1 1x101 1x102 1x103 1x104 1x105 1x106 1x107 1x108
X-ray/neutron techniques A ruler
Conventional microscopy
Electron microscopy
Dynamic Light Scattering
Particle Tracking techniques
Applications Of DLS
• We measure Hydrodynamic Size of nanoparticle ,protein and
biomaterial
• We can also study stability of nanoparticles as function of time
• Good for detecting the aggregation of the particles
Other Then these
• Required small volume of sample
• Complete recovery of sample after measurement
• Sample preparation is not required for the measurement
Brownian motion
• Brownian motion is the fundamental of this instrument
• Brownian motion of the particle is random motion due to the
bombardment by the solvent molecule surround them.
• Brownian motion of the particles are related to size.
• It describes the way in which very small particles move in fluid
suspension
Stoke’s Einstein Equation: Relating
particle size to particle motion
• This
random motion is modeled by the Stokes-Einstein
equation.
d=
• d is the hydrodynamic radius of the particle ,the diameter of
the sphere that has same diffusion coefficient as the particle.
• Temperature of the measurement must be stable ,as the
viscosity of the liquid.
Hydrodynamic radius
• “The size of a hypothetical hard sphere that diffuses in the
same fashion as that of the particle being measured”
How DLS Works
• In DLS we measured the speed at which the particles are
diffusing due to Brownian motion.
• Speed of diffusion is measured by measuring the rate at which
the intensity of the scattered light fluctuates.
• Small particles causes the intensity to more fluctuate than
larger.
• It measure the diffusion coefficient by using correlation
function.
How these fluctuation in scattered
light arises ?
For the particle in Brownian
motion a speckle pattern
is observed where the
position of each speckle is
seen to be in constant
motion Because the phase
addition from moving
particle is constantly
evolving and forming new
pattern.
Speckle Pattern for a
sample containing
Stationary Particle
Experimental setup of the DLS
LASER
Detector
Attenuator
173
Digital
signal
Processer,
Sample
90 Correlator
cell
Detector Software for
Scattered light data analyzer
DLS signal
Obtained optical signal shows random change due to random change
in the position of the particle.
The “ noise “ is actually the particle motion and will be used to measure the particle
size.
Correlation function
• A correlation function is statistical correlation between
random variables at two different points in space or time,
usually as a function of the spatial or temporal distance
between the points.
• Within the correlation curve all of the information regarding
the diffusion of particles within the sample being measured.
• Correlator construct correlation function G(τ), of the scattered
intensity
G(τ)=<I(t).I(t+τ) ,τ is delay time
If the intensity at time t is
Cont. compared with the intensity at
time t+δt, there will be a strong
correlation between the two
signals.
Correlation of a signal arriving
from random source will decrease
with time.
If the particle will large the signal
will changes slowly and correlation
will sustain for long time.
Correlogram
Typical intensity fluctuations
for large and small particles and
Corresponding Correlogram
How the correlator works
• It is a signal comparator. It is design to measure the similarity
between two signals or one signals with itself with varying
time.
• The correlogram give many information the time at which the
correlation starts significantly decay is an indication of
mean size of the sample
• The steeper the curve the more mono disperse the sample is.
More extended the decay becomes the greater the
polydispesity.
Correlation function for
monodispresed Particle
•
For monodispersed particle in Brownian motion , the correlation
[G] function is an exponential decaying function of decay time τ
G(τ) =A[1+Bexp(-2Г τ)]
Delay constant (Г) is related to the diffusion coefficient of the
particle ,D
Г=
q is the scattering vector which is given by
and q=
The correlation function for
polydisperse particle
• G(τ) =A[1+Bexp(2(-Г’ τ+ẚ/2+----)]
Г’ is delay constant
Г’=
D’ is the diffusion coefficient
Polydispersity Index =
• measure the distribution of size in our sample.
• for monodisperse it is about 0.01-0.05 but the value
greater than 0.7 indicate that sample has a very broad
size distribution.
Comparative techniques
DLS Particle Tracking Transmission Electron
microscopy
Size range (depending on
Size range: 0.3nm to 10µm Size range: 10nm to 2 µm resolution from sample): 1 nm
up to a few µm
Size, polydispersity, and zeta Size, diffusion coefficient, and
Size and morphology
potential concentration
Detects particles of all sizes Detects particles of all sizes
Biased towards larger particles (within the size range)
Quick and easy to run samples Easy to use, but more labour Can be time consuming and
intensive expensive
Highly reproducible data, but
Highly reproducible data Requires lots of images to gain
(assuming sample is good and requires more runs, as only a an accurate representation of
small volume of sample is
without contamination) measured particles
Generates images. Needs
Data produced can be Data produced is dependent on
misleading operator settings specialist expertees to gain
statistical data
High start-up cost, low High start-up cost, low running High start-up cost and running
running cost cost cost
Limitations of DLS
• We measure the hydrodynamic radius of the particle, not able
to measure the actual size of the particle
• The particles having size greater than 1000nm are not
measured by this method
• Size of Solid particles are not measured by DLS
Summary
• Good technique for gaining a particle
size/dispersity/zeta potential
• High output and easy to use
• Can measure thermoresponsive systems effectively
• Biased towards larger particles
• Struggles with polydispersed samples
• Data analysis and quality control can be complicated
• Not absolute, and should be used in conjunction to
other techniques
References
• DLS technical note www.malvern.co.uk
• http://www.wyatt.eu/
• Dynamic Light Scattering for Nanoparticle Size Analysis -
HORIBA
Thank you for your attention
