High-Concentration Submicron Particle Size Distribution

by Dynamic Light Scattering

Power spectrum development with heterodyne technology advances

biotechnology and nanotechnology measurements.

M. N. Trainer and P. J. Freud

Application Note

SL-AN-05 RevA

Provided By:
Microtrac, Inc.
Particle Size Measuring Instrumentation

SL-AN-5 Rev A 1 05/30/03

DYNAMIC LIGHT SCATTERING (DLS) is a well-established technique for measuring particle size over
the size range of a few nanometers to a few microns; however, at high sample concentrations severe
limitations are placed on the DLS measurement. This paper discusses the causes of the high concentra-
tion limitations, the means of overcoming the limitations, and the results of measurements at high
concentrations.

DLS determines particle size from the analysis of the Brownian motion of suspended particles. Light scat-
tered from a moving particle has a Doppler light frequency shift imparted to it. Scattering from a group of
particles will have a distribution of shifts from the randomly moving particles. Figure 1 illustrates two
measurement configurations that can be employed to extract the Brownian motion information from the
frequency shifted, scattered light. Homodyne detection shown on the left extracts the shifts by the
interference between the light scattered from each particle with the light scattered by the rest of the
particles. The interference or self-referencing removes the high optical frequency, leaving the lower shift
frequencies. This is the conventional photon correlation spectroscopy (PCS) geometry, usually measured
at a 90° scattering angle.

HOMODYNE HETERODYNE
PARTICLES PARTICLES
INCIDENT LIGHT INCIDENT LIGHT

is io
is is

DETECTOR DETECTOR

2 2γ γ
P(ω) = < is> P(ω) = io < i >
2
ω + (2γ) 2 s
ω2 + (γ)2

kT q = 4 π sin( θ / 2)
γ = D q2 D
6πηa λ
Figure 1. Comparison of homodyne and heterodyne configurations used for dynamic
light scattering. Io represents the reflected, reference laser. IS represents the scattered
light from particles. P(ω) represents the distribution of frequencies related to the
distribution of particle size

In the heterodyne measurement, shown on the right, a portion of the incident light is diverted from the
scattering particles and then mixed with the scattered light. The unshifted diverted light acts as a
reference or local oscillator (hence heterodyne) for the shifted scattered light from each particle.

In both cases, the interference signal frequencies are indicative of the Doppler frequency shifts (due to
particles' Brownian motion) and are the basis of the particle sizing measurement of DLS. The expressions
shown in Figure 1, P(ω), represent the power spectrum of the distributions of frequencies. These are
Lorentzian functions of angular frequencies, ω, for both homodyne and heterodyne detection. The particle
size is determined by the analysis of these power spectra. The constant, γ, appears as a characteristic

SL-AN-5 Rev A 2 05/30/03

frequency of the response and is a function of the wavelength, λ, scatter angle, θ, and the diffusion
coefficient, D. The diffusion coefficient,D, depends on the temperature, T, suspending medium viscosity,
η, and particle radius, a.

Note that there are two differences between the power spectra for the two cases. First, the homodyne
detection power spectrum depends upon twice the characteristic frequency, γ, while the heterodyne
detection power spectrum depends upon just ω. Second, the homodyne power is proportional to the
scattered light intensity squared, (Is)2, while the heterodyne power is proportional to the product of the
scattered intensity and reference intensity, (Is) (Io).

When measuring in the homodyne mode, only the homodyne expression applies. Conversely, when
measuring in the heterodyne mode, a mix of the two modes will apply. The effect of mixing the two modes
together will be to mix power spectrum of characteristic frequency, γ, and power spectrum of
characteristic frequency, 2γ, with a resulting ambiguity of particle size. The mixing will depend upon the
relative magnitude of each mode. By providing a large reference intensity, the heterodyne mode can be
made large enough to dominate the mixture of the two.

The Microtrac NANOTRAC operates primarily with heterodyne detection. Figure 2 is a diagram of the
NANOTRAC measurement system. A magnified view shows the region of optical interaction with the
suspended particles. The analyzer has an optical waveguide immersed in the suspension. The 50-µm
diameter waveguide delivers the input beam to the sample and collects the backscattered light from a
region within 100 urn of the waveguide-medium interface. The Fresnel reflection at the interface between
the waveguide and the medium is mixed with the backscattered light. The reflection is unshifted in
frequency and provides the local oscillator for heterodyne detection.

FIBER OPTIC
CONNECTION
TO 100 m PARTICLE
STAINLESS STEEL SUSPENSION
OPTICAL SPLITTER
LASER OPTICAL PROBE

PHOTO

100 µ
Scattered

Laser
Input Reflected
SUSPENDED
PARTICLES
WAVEGUIDE

Figure 2. Diagram of the implementation of the Controlled Reference Method used in the
Nanotrac. The term relates to the use of a reflected beam as a reference oscillator to
isolate the scattered, frequency-shifted light. The waveguide includes the optical splitter,
fiber optic connector and the enclosed stainless steel probe.

Two important features should be noted. First, the high-intensity reflected reference allows the
heterodyne component to dominate the power spectrum. Second, the very short pathlength, the 100-µm
that the scatter signal has to travel, minimizes multiple optical scattering effects even at high particle
concentration.

Also shown in Figure 2 is the balance of the measurement system for the analyzer. The tip of a three-port
surface waveguide is immersed in a sample cell containing the suspended particles. The directional “Y”
optical splitter delivers the input from the laser diode to the tip and returns the scatter and the reflected
beams to the photodetector. The power spectrum of the interference signal is calculated with dedicated

SL-AN-5 Rev A 3 05/30/03

high-speed FFT (fast Fourier transform) digital signal processor hardware. The power spectrum is then
inverted to give the particle size distribution.

In considering the effects of high concentration on the dynamic light scattering particle size measurement,
a number of mechanisms that alter the apparent particle size can be considered. The possibilities listed in
Figure 3, are divided into two groups: those which increase the apparent size and those which decrease
the apparent size.

Increase Apparent Size Decrease Apparent Size

Van Der Waals Forces Interparticle Repulsive Forces
Agglomeration Multiple Scattering
Other Interparticle Attractive Forces Homodyne (self-beating) Mixing

Figure 3. Mechanisms for concentration depence of apparent size in Dynamic Light Scattering measurements.

Interparticle attractive forces such as Van Der Waals result in a net apparent size increase with
concentration. Agglomeration as concentration increases would also result in an apparent size increase.
The occurrence of agglomeration can usually be established since it tends to be irreversible.

Repulsive forces result in an apparent size decrease as concentration increases. Two instrumentation
effects would cause apparent size decreases as concentration increases. Multiple scattering results from

Change of Measured Size

as Sample Concentration Changes
0.200u Polystyrene
(Theoretical relative size = 1.0)
1.2
1.0
0.8
Relative
Size 0.6
0.4
0.2
0
1 10 100 1000 10,000 100,000
Concentration, ppm

Nanotrac 150 PCS Competitor

0.705 µ Polystyrene
1.2
1.0
0.8
Relative
Size 0.6
0.4
0.2
0
1 10 100 1000
Concentration, ppm
Figure 4. Concentration dependence of apparent size for polystyrene samples

SL-AN-5 Rev A 4 05/30/03

light interacting with particles multiple times. The scattering and re-scattering causes multiple frequency
shifts at different angles. The net effect is an apparent size reduction. A second effect occurs if
heterodyne detection is used. An interference from the homodyne component with its doubled
characteristic frequency will be present and will cause a decrease in apparent size. The effect will depend
on how large the homodyne component is relative to the heterodyne component.

The concentration dependence of the apparent size of a number of polystyrene sphere samples has been
determined using the Microtrac NANOTRAC and a PCS instrument. The comparison is shown in Figure
4. The PCS instrument utilizes 90° scattering, homodyne detection, and a cuvette sample cell with a
scatter pathlength of approximately 0.5 cm. The effects of multiple scattering are evident for the PCS long
path measurement. A large decrease in apparent size is seen at relatively dilute samples. The short path
NANOTRAC measurement shows no change in apparent size for the 200-nm polystyrene sample in
Figure 4. For larger 460-nm and 705-nm polystyrene, the PCS measurement again shows the effects of
multiple scattering at its upper concentration limit, while the NANOTRAC continues to show a
concentration-independent size.

Another advantage of the heterodyne technique is illustrated in Figure 4 for the 460-nm and 705-nm
measurements at the low concentration end. The improved signal-to-noise ratio of the heterodyne
measurement allows the NANOTRAC to measure at lower concentrations, while the PCS instrument fails
to measure at the same concentrations due to the inherently lower signal level of the homodyne
measurement.

The comparisons shown in Figure 4 illustrate that a short-path method is required for high-concentration
dynamic light scattering size measurements in order to avoid multiple scattering and the resulting size
errors. At the highest concentration range, only the short-path method of the NANOTRAC measurement
is considered. With the concentration extended to 30% for 200-nm polystyrene, Figure 5 illustrates that
the analyzer continues to measure concentration-independent size up to a few percent concentration,
while above the few percent concentration range, the apparent size increases, reaching a value 30%
higher than that of the nominal size.

1.4

1.2

1.0
Relative 0.8
Size
0.6

0.4

0.2

0
.01 0.1 1.0 10 100

Concentration, ppm
Figure 5. Concentration dependence of apparent size for 200nm
polystyrene in water at high concentrations.

To determine the sources of the high-concentration size dependence, one additional aspect of the
measurement system needs to be considered. Although the immersed waveguide measurement
emphasizes the heterodyne contribution in the measured power spectrum, a homodyne component is
mixed in that will shift the apparent size to a smaller value. The question is, how large is the shift? The
probe medium reflectance determines the magnitude of the reflected reference and the magnitude of the

SL-AN-5 Rev A 5 05/30/03

heterodyne component. The reflectance depends upon the difference between the index of refraction of
the waveguide and of the suspending medium. The dependence is shown in Figure 6. For a silica
waveguide, i.e., fiber optic, with an index of 1.46, the reflectance decreases as the medium index
approaches the silica index. When the two indices are matched, the reflectance is zero and the
NANOTRAC measurement would be pure homodyne with no heterodyne component. The apparent size
would be half the nominal size. (Carbon tetrachloride, toluene, and trichloroethylene are examples of
suspending media that almost match a silica optical waveguide.)

0.025

0.020

PROBE 0.015 Sapphire

REFLECTIVITY
0.010

0.005 Silica 50% 200nm

polystyrene
0.000
1.30 1.35 1.40 1.45 1.50

SUSPENDING MEDIUM INDEX

Figure 6. Reflectance dependence on suspending medium index

for silica and sapphire waveguide probes. Equivalent points for 50%
concentration 200nm polystyrene in water indicated by arrows.

A probe has been developed for use by the analyzer that avoids this problem. Sapphire, with an index
of1.76, is employed as a tip on the silica waveguide. The sapphire tip interfaces with the suspension and
eliminates the concern of index matching. Figure 6 illustrates the reflectance curve versus medium index
for sapphire.

Even if the suspending medium is water, the effective index of the suspension can take on a high value at
high particle concentration. The suspension takes on an index that is approximated by a volume average
between the particle index and the suspending medium index. The lower plot of Figure 6 illustrates the
case of polystyrene, with an index of 1.59; mixed with water, with an index of 1.33. The net index results
in index matching with a silica interface at concentrations of 50%, while the sapphire-tipped probe
maintains a high value of reflectance even at the highest concentrations of polystyrene.

With the sapphire-tipped probe, the particle size was measured for 200-nm polystyrene up to 30%
concentration. As shown in Figure 7, the measured size approaches a linear dependence on
concentration in this concentration range. The homodyne mix correction, calculated for the decrease in

1.
200 NM
POLYSTYRENE
MEASURED

1.

RELATIVE
DIAMETER CALCULATED
HOMODYNE MIX
0. CORRECTION

0.
0 1 2 3
VOLUME CONCENTRATION,
Figure 7. Concentration dependence of apparent size for
200nm polystyrene.

SL-AN-5 Rev A 6 05/30/03

probe reflectance with concentration, would produce a net size decrease as shown in this figure. The
cause of the difference between the measured size and the corrected size is the concentration
dependence of interparticle interaction, an inherent property of the suspension.

Concentration-dependence measurements were made with the analyzer on three sizes of polystyrene.
The relative size, that is, the measured size relative to the low concentration size, is plotted in Figure 8.
The three sizes of polystyrene fit a common linear dependence on concentration indicating a common
interparticle interaction for these similar particle suspensions.

1.25

1.20

RELATIVE 1.15
APPARENT
SIZE
1.10
140
204
1.05 394
LINEAR FIT
1.00
0 5 10 15 20
VOLUME CONCENTRATION, %

Figure 8. Concentration dependence of apparent size for three

sizes of polystyrene (140nm, 200nm, and 394 nm).

In summary, the authors have shown that high concentration particle size measurements can be made
using dynamic light scattering. The Microtrac NANOTRAC incorporates a direct-immersion waveguide
with its inherent short optical pathlength to minimize multiple scattering errors. The sapphire probe with
high tip reflectance provides a high heterodyne reference intensity, minimizing the homodyne mixing
error. Interparticle interaction is the principal mechanism for size concentration dependence in the very
high-concentration region, with three different polystyrene samples showing a common concentration
dependence.

SL-AN-5 Rev A 7 05/30/03