TECHNICAL NOTE

Polarization-resolved microscopy
and spectroscopy solutions

This Technical Note highlights Oxford

Instruments WITec’s Raman imaging
microscopy and spectroscopy solutions
for exploring polarization-dependent
1.4
1.2

effects.
Intensity [a.u.]

1.0
0.8
0.6
0.4
0.2
0

300

Po
lariz
ati 200
ona
ng 3000
le
[°]
100 2000
l. cm ]
-1

shift [re
1000 Raman
0
TECHNICAL NOTE

Polarization-resolved spectroscopy and microscopy

Polarization is a property of light that can In Raman spectroscopy, precise control In this Technical Note, we illustrate
be exploited with various microscopy of the polarization of both the excitation the potential of polarization-resolved
techniques to obtain valuable insights into and scattered light offers an effective spectroscopy and imaging on samples
a sample’s characteristics, including its means for determining the symmetry from the fields of geology, polymers,
structural orientation, optical anisotropy and directionality of vibrational modes. biomineralization studies and 2D
and chirality. In highly ordered substances such as materials research.
Polarized white light microscopy is polymers, 2D materials and bulk crystals,
widely used in geosciences to analyze polarization-dependent experiments
the composition and optical properties of illuminate structural properties including
crystalline minerals. Further applications anisotropy.
include the quantification and structural
analysis of organic specimens, polymers
and fibers.
Polarization-dependent second-harmo-
nic generation (SHG) measurements are
a powerful approach for characterizing
crystal orientation and strain fields in
samples such as 2D atomic crystals,
which are of great interest to the
semiconductor industry (for further
information, see the WITec Application
Note on Correlative Imaging of MoS2 [1]).
Polarization-resolved photolumi-
nescence spectroscopy (PL) reveals
detailed information on electronic
band structure and exciton dynamics in Figure 1: Parallel polarized white light transmission image (left panel) and Raman image
semiconducting materials. made using TrueComponent Analysis (right panel) of a 25µm thin section of black mica
revealing different crystal orientations.

Microscope setup with polarization control

All measurements presented here were scattered light to be studied. By rotating
performed with Oxford Instruments the analyzer, the directional emission
WITec Raman microscopes equipped pattern of a vibrational mode can be
with integrated polarization modules. probed. Both polarizer and analyzer are
For polarization-resolved white light independently rotatable in alpha300
imaging, two independent polarization systems, offering complete freedom in
filters were introduced to the optical selecting polarization configurations. The
beam path: the first (polarizer) is optional motorization of the analyzer and
positioned behind the unpolarized white polarizer modules enables the recording
light source, the second (analyzer) of automated polarization-dependent
after the sample in the detection beam imaging series (see example abalone
path. For all laser-based polarization- shell).
resolved spectroscopic measurements There is also the option of placing an
including Raman spectroscopy, SHG additional λ/2 or λ/4 wave plate in the
and photoluminescence spectroscopy, combined excitation and detection
dedicated polarizer and analyzer beam path. The λ/4 wave plate enables
modules were used (Figure 2): Here, the dedicated studies of the helicity of
polarizer consists of a freely rotatable the scattered light, which is useful for
half-wave plate that precisely orients investigating the sample‘s out-of-plane
the linearly polarized excitation beam’s orientation [2].
polarization on the sample. Optionally, a
quarter wave plate can facilitate left- or
right-handed circularly polarized light.
The analyzer module in the detection Figure 2: Polarizer and analyzer modules
beam path again allows for the selection in a WITec alpha300 R system
of a specific linear polarization state of the
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Info Box: Polarization-resolved Raman spectroscopy

Raman spectroscopy that makes use of polarization effects relies on the electromagnetic nature of light, as polarization corresponds
to the direction along which the electric field is oscillating. The wave equation (derived from Maxwell’s equations) dictates that
the magnetic and electric fields of the electromagnetic wave always oscillate perpendicular to its propagation direction. While
unpolarized light is a superposition of waves with their electric fields oscillating in random directions, the electric fields of all waves
are oriented in parallel for polarized light. Dependent on the molecular symmetry of the sample, the vibrational modes visible in
the Raman spectra can be sensitive to the polarization angle of the excitation light. This reflects in a vibrational mode’s Raman
signal strength varying with the angle of polarization of the incident light, as visible in many in-plane anisotropic samples. At the
same time, unoriented or isotropic crystal/molecular structures show no pronounced polarization dependence in their individual
Raman peak intensities. Consequently, polarization-resolved Raman spectroscopy can provide much more profound information
on the studied sample such as its structure and orientation of different domains.

Excitation path Detection path

Figure 3: Schematic image of polarization control components.

Excitation path: The coherent, linearly polarized incident light is emitted from a laser with its electric field oscillating perpendicular to the propa-
gation direction. The linear polarization state can be rotated to the desired orientation using a half-wave (λ/2) plate in the polarizer module. A
quarter-wave (λ/4) plate may be introduced to produce elliptically or circularly polarized light.
Detection path: Using a polarization filter, i.e., the analyzer in the beam path before the detector, the polarization state of the detected light can
be selected.
TECHNICAL NOTE

Visualizing the molecular processes in polyethylene stretching

Introduction Experimental details was used for the stress measurements
Polyethylene (PE) is a synthetic polymer The experiments were carried out at of the polyethylene foil. Here, the
with the chemical formula (CH2)n, which is Swiss Federal Laboratories for Materials material was stretched from 15 mm in
one of the most commonly used materials Science and Technology (EMPA). its unstressed state up to 40 mm when
for plastic packaging. Its molecular fiber Automated polarization series were fully stretched (Figure 4) and polarization
structure and the possibility of introducing obtained over the full polarization rotation series were obtained at 10, 15, 20 and
crosslinks between the fibers allow for (360°) of the electric field of the laser 25 mm of stretching. All measurements
the creation of plastics with different light with one spectrum being recorded were performed with a 532 nm excitation
physical properties that are useful for a at every degree of rotation. A Linkam laser and a 50x/0.55 LWD objective.
wide range of applications. Commercial modular force stage (Linkam MF-stage)
PE products are sorted into two major
groups: High-density PE (HDPE), including A B
more rigid materials and those that
appear more opaque, and low-density
PE (LDPE), which provides high flexibility
and is more transparent. As such, LDPE
is commonly used as plastic wraps for
food packaging to preserve and protect
the food from chemical, biological and
physical factors in the environment. In
the following experiments, polarization-
dependent Raman measurements were
used to investigate the effects of plastic
foil stretching on the molecular PE fiber
orientation.

Figure 4: Picture of the experimental setup.

PE sample fixed within the Linkam modular force stage under the Raman microscope. (A) Ma-
terial unstretched (15 mm) (B) Material fully stretched (40 mm).

A B
5
5
1 2 3 4
4.0 4.0 1 2 3 4
[log(10) CCD cts]

[log(10) CCD cts]

3.8 3.8
3.6 3.6
Intensity

Intensity

3.4 3.4
3.2 3.2
3.0 3.0
2.8 2.8
2.6 2.6

300 300
Po Po
l ar l ar
iza 200 iza 200
tio tio
n n
an 100
an 100
gl 3000 gl 3000
e e
[°] 2000 [°] 2000
1000 -1 ] 1000
]-1
shift [rel. cm shift [rel. cm
0 0
0
Raman 0
Raman
Intensity [ CCD cts]

2000

1500

1000

Figure 5: 3D heatmap of polarization-resolved Raman spectra in PE foil stretching. 500

(A) Unstretched polyethylene foil. (B) Fully stretched (ΔL/L0 = 5/3) state, inlet shows mag- Po
300
l ar
nification of the fingerprint area (800 - 1600 rel. cm-1). Labelled Raman peaks: (1) νas(C-C) iza 200
tio
n
at 1063 rel. cm-1; (2) νs(C-C) at 1130 rel. cm-1; (3) τ(CH2) at 1297 rel. cm-1; (4) δ(CH2) at 1417, an
gl
e
100
1400
[°]
1441 and 1464 rel. cm-1; (5) νas(CH2) and νs(CH2) at 2849 and 2883 rel. cm-1 [3-5].
1200
1000 -1 ]
0
ft [rel. cm
Raman shi
800
WITec Wissenschaftliche Instrumente und Technologie GmbH
Lise-Meitner-Str. 6, D-89081 Ulm, Germany
Tel. +49 (0) 731 140 700, Fax +49 (0) 731 140 70200
info@WITec.de, https://raman.oxinst.com

Results Based on the obtained data and changing Conclusion

The polarization-resolved analysis Raman intensities at different polarization The presented experiment is an example
revealed a characteristic Raman spectral angles, the orientation direction of the of how a materials‘ structural orientation
pattern for PE with five major peaks, which PE fibers within the plastic foil can be can be determined using polarization-
were independent of the polarisation deduced. While molecular bonds parallel dependent Raman spectroscopy. As the
direction of light in the unstretched to the polarization direction of the laser polarization orientation of the incident
material (Figure 5A). In contrast, the light cause high Raman band intensities, laser light was a known factor in the
Raman modes for the symmetrical C-C decreased Raman signals are detected measurements, the macroscopic PE fiber
stretching νs(C-C) (1130 rel. cm-1), CH2 for perpendicularly oriented bonds. orientation in the plastic wrap was easily
bending δ(CH2) (1417, 1441 and 1464 Hence, for the investigated sample, the established. The method is especially
rel. cm-1) and C-H stretching ν(C-H) (2849 following interpretation can be made: useful for investigating the same sample
and 2883 rel. cm-1) showed polarization- the isotropic orientation of the PE fibers in different conditions, such as the
dependent intensity changes in the fully in unstretched condition is modified when different measures of stretching shown
stretched condition (Figure 5B). applying a pulling force to the material. here, due to its nondestructive nature and
In the following, the νs(C-C) and the C-H In the stretched condition, the PE fibers as it does not require additional sample
stretching modes, both being polarization show a highly oriented (anisotropic) preparation.
dependent but in opposite angular distribution, with the longitudinal axis of
orientations, were investigated in more the fibers in line with the polarization of
Orientation of the Polarization direction
detail during the PE stretching process. the laser light at 90° (Figure 7). The C-H stretched PE fiber of the laser light at 90°
For data evaluation, their spectra were stretching bonds align in the orthogonal
normalized to the Raman band at 1063 orientation to this axis with increasing
rel. cm-1 of asymmetric C-C stretching, as pulling force. H C H 90°
no strong polarization dependence of the
Raman signal was observed here in both A νs(C-C) (1130 rel. cm-1) H C H
180° 0°
the unstretched and stretched condition. 90

The respective polar plots are shown in 120 60 H C H

Figure 6. 270°
In its relaxed state, the PE foil shows a 150 30
H C H
homogenous Raman intensity for all
polarization angles for both investigated
α [°] Figure 7: Scheme of the stretched PE fibers.
spectral regions (Figure 6 – yellow color).
180 0
This can be explained by an isotropic Orientation in relation to the polarization
distribution of the fibers, exhibiting no direction of the exciting light.
predominant direction in the material.
Stretching the foil with the Linkam MF- 210 330

stage by 10 mm from its initial 15 mm

(unstretched) to 25 mm (ΔL/L0 = 2/3) 240 300
results in differences in the Raman 270

signals with respect to the polarization

of the incident light (Figure 6 – orange B ν(C-H) (2800 -3000 rel. cm-1)
90
color). The C-C stretching shows intensity
120 60
minima for 0° and 180° polarization
angles and maxima at 90° and 270°. The
intensity pattern of the CH-stretching 150 30

mode follows the inverse orientations

(maxima at 0° and 180° and minima at
90° and 270°, respectively). Stretching α [°]
180 0
the foil even further by 25 mm (ΔL/L0 =
5/3) to a total length of 40 mm enhances
these intensity differences between the Figure 6: Polar plots of PE foil within
330
polarization angles (Figure 6 – red, purple 210
stretching.
and blue). (A) Symmetrical C-C stretching band at
240 300 1130 rel. cm-1. (B) C-H stretching region at
270 2800 - 3000 rel. cm-1. The dots represent
the experimental data, the line shows the
ΔL/L0 = 0 ΔL/L0 = 2/3 fit. Stretching color coded from unstressed
ΔL/L0 = 3/3 ΔL/L0 = 4/3 ΔL/L0 = 5/3 (yellow) to ΔL/L0 = 5/3 (blue).
TECHNICAL NOTE

Polarization-resolved Raman spectroscopy of 2D materials

Introduction 2D semiconductor MoO3 single-coordinated (O(1)) along [010],
Ever since the fabrication of monolayer Here, we focus on the in-plane anisotropic double-coordinated (O(2)) along [100],
graphene in 2004, the field of 2D van-der Waals semiconductor molyb- and tri-coordinated (O(3)) along the [001]
materials research has been motivated denum trioxide (αα-MoO3) with a direct direction. The different Mo-O bonds along
by their intriguing optical, electronic, and electronic bandgap of ~3 eV (410 nm). the crystallographic axes eventually give
mechanical properties. Recently, in-plane MoO3 exhibits an orthorhombic crystal rise to a plethora of vibrational modes,
anisotropic 2D semiconductors have structure (space group Pnma) with many of which are Raman- or IR-active,
become subject of particular interest due the lattice constants a = 0.396 nm, b with the latter supporting in-plane
to their properties being not only restricted = 1.382 nm, and c = 0.373 nm, where hyperbolic phonon polaritons [6].
in 2D, but also exhibiting an inherent in- the van-der-Waals stacking direction
plane directionality. Naturally, this young is along [010] (Figure 8). The vdW layer Experimental details
scientific field requires fast, high-resolution consists of a double layer of distorted The investigated 25 × 4 µm² MoO3 flake
tools for material characterization. To this octahedra, wherein molybdenum and depicted in Figure 9 was exfoliated
end, the benefits polarization-resolved oxygen atoms are covalently bound. onto a SiO2/Si substrate with the
Raman spectroscopy can provide for 2D The in-plane anisotropy stems from the MoO3 bulk crystal grown using the
materials research and engineering are arrangement of three classes of oxygen Bridgman technique. As the material
illustrated in the following. atoms with different oxidation states: typically breaks parallel to its in-plane
crystallographic axes, the flakes come
Oxygen in a rectangular shape. The polarization
Molybdenum geometry is illustrated in the inset
(Figure 9). Here, the polarization angle
α of incident light is defined against the
microscope’s x-axis with the analyzer in
the detection beam path always parallel
to the incident polarization direction. All
measurements were performed using
a 100x/0.9NA objective and 1.0 mW of
[001]

[010]

[010]

power at 532 nm excitation wavelength.

Polarization-resolved Raman
[100] [001] [100] spectra
[010] [100] [001]
Raman spectra of the MoO3 flake with
Figure 8: Crystal structure of α-MoO3. incident and detected polarization
parallel to the [100] (black) and [001]
Crystal structure shown in three different orientations. Black box indicates the unit cell with the
parameters: a = 0.396 nm, b = 1.382 nm, c= 0.373 nm. direction (red) are shown in Figure 10A:
The Raman spectra reveal several sharp
Representation generated with Crystal Toolkit [7].
peaks in the 0 to 1100 rel. cm-1 range,
many of which exhibit a pronounced
Si Substrate polarization dependence. For instance,
the Raman mode at the Raman shift of
160 rel. cm-1 (highlighted blue) has its
maximum when excited along the [001]
direction, whereas the Raman mode at
]
[100

MoO3 flake 820 rel. cm-1 (yellow) shows the opposite

behavior. Moreover, some Raman active
modes are barely visible when excited
[001
] parallel to an in-plane crystal axis, for
instance at 284 and 667 rel. cm-1 (shaded
y red and green), which can be attributed
Figure 9: Optical image of MoO3 sample. to the nature of the vibrational mode.
α Note that the first and second order Si
Investigated 25 × 4 µm² MoO3 flake with the x
in-plane crystal axes indicated. The inset peaks stemming from the substrate are
depicts the layout of the polarization in the visible due to the finite thickness of the
5 µm polarization
experiment. MoO3 flake.
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Tel. +49 (0) 731 140 700, Fax +49 (0) 731 140 70200
info@WITec.de, https://raman.oxinst.com

Polarization-resolved Raman A
intensities
pol.. || [100]
pol [100]
In order to obtain additional information
pol.. || [001]
pol [001]

[a. u.]
about the four selected modes, a

[a.u.]
measurement series was performed in

intensity
which the angle of incident polarization

Intensity
α together with the analyzer was rotated
from 0° to 360° and a spectrum recorded

Raman
every 5°. This measurement layout (Si)
consisting of a stationary sample and
controlled polarization is particularly
advantageous for 2D material
investigations as the position of the Raman shift -1
Raman shift[rel.
[rel.cmcm
] -1]
excitation on the sample doesn’t change,
allowing for automated measurements B Ag-mode (160 rel. cm-1) B2g-mode (284 rel. cm-1)
(the presented series took 2.5 minutes
to record). The results for the four modes
are displayed in Figure 10B in individual
polar plots, with the respective integrated
Raman intensity on the r-axis. Here, the
experimental data are represented by α [°] α [°]
the data points and the black curves
correspond to fits according to [8].
• Ag-mode at 160 rel. cm-1: [001
]
Two-lobed mode with its maxima orien-
ted parallel to the [001] direction.
• B2g-mode at 284 rel. cm-1:
Four-lobed mode with its maxima 45°
off-axis.
B2g, B3g-mode (667 rel. cm-1) Ag, B1g-mode (820 rel. cm-1)
• B2g, B3g-mode at 667 rel. cm-1:
Four-lobed mode with its maxima 45°
off-axis.
• Ag, B1g -mode at 820 rel. cm-1:
Two-lobed mode with its maxima ori- ]
[100

ented parallel to the [100] direction (Ag α [°] α [°]

component) superposed by a weaker
four-lobed shape that has its maxima
along the main crystal axes (B1g com-
ponent).
Overall, the fits nicely match the data and
the shapes of the modes correspond with
data found in literature [9].

Conclusion Figure 10: Polarization-dependent Raman spectra of MoO3.

(A) Raman spectra of MoO3 with both incident and detected polarization along the [100] (black)
Using the example of molybdenum
and [001] directions (red). (B) Integrated Raman intensity of different vibrational modes as a
trioxide, we demonstrated the great
function of angle of polarization α. The polar plots represent the experimental data (datapoints)
potential polarization-resolved Raman and their corresponding fits (black lines).
spectroscopy offers to the field of
2D materials: fast, nondestructive
assignment of crystallographic axes and
determination of the type of vibrational
modes. This is in addition to the standard
properties accessible with Raman
spectroscopy, such as information about
crystal quality, stress/strain and material
composition.
TECHNICAL NOTE

Polarization-dependent Raman imaging of an abalone shell

Introduction Sample preparation A
In the search for materials with great Samples were cut from an abalone shell
stiffness, strength, and toughness, (Haliotis rufescens) using a diamond saw.
mollusk shells serve as a source of Nacre samples (~ 20 × 5 × 5 mm3) were
inspiration for the design of advanced embedded in epoxy resin with the c-axis
functional materials [10]. Over the years parallel to the surface and manually
biominerals have been refined by natural polished in dry conditions with a series of
selection, leading to the unique forms SiC sandpapers of decreasing grain size
and functions we observe today. For to reach a surface roughness of a few
studying their exceptional structure, hundred nanometers.
polarization-resolved Raman imaging is
a powerful tool that can reveal biomineral RISE measurements on nacre
composition and crystal orientation. In this study, correlative Raman and
SEM imaging was used to examine the
Abalone (Haliotis rufescens) shell aragonite phase of the nacre, where
An abalone shell consists of polymorphs the platelets form a brick-and-mortar
of CaCO3 [11]. The outer part of the shell structure perpendicular to the growth B
is formed of calcite with a trigonal R3c direction (Figure 12A).
crystal form. The translucent inner part Imaging was performed with a Raman
called nacre is built up from platelets Imaging and Scanning Electron (RISE)
of aragonite. The orthorhombic crystal Microscope in low vacuum mode at 15
structure of aragonite with a unit cell Pa and an acceleration voltage of 15 kV.
(Pnma space group) with the lattice Raman data were collected from an area
Intensity [a.u.]
constants: a = 0.495 nm, b = 0.796 nm of 20 x 15 µm2 (100 x 75 Raman spectra,
and c = 0.573 nm is shown in Figure 11. 0.2 s iintegration time per spectrum),
using a 532 nm excitation laser at 10
mW. For the polarization-dependent
Carbon
Calcium
Raman measurements, the incident laser
Oxygen light was oriented in the growth direction
of the aragonite platelets (horizontal
direction in Figure 12A). On the detection
side, all of the Raman scattered light was Raman shift [rel. cm-1]
collected. C
The Raman spectra acquired from the
nacre show a characteristic pattern for
Figure 11: Crystal structure of aragonite. aragonite (Figure 12B). Raman bands
Unit cell (black box) with the dimensions a = at low wavenumbers originate from the
0.495 nm, b = 0.796 nm and c= 0.573 nm. CO3 lattice vibration modes such as the
translational mode and wobbling (153
rel. cm-1 and 206 rel. cm-1, respectively).
The strong peak at 1085 rel. cm-1
corresponds to CO3 stretching [12]. When
the acquired Raman image is overlaid
onto the SEM image (Figure 12C) the
platelet‘s co-orientation along the growth
direction is revealed. This corresponds to
the c-direction in the schematic in Figure
11.
Figure 12: RISE microscopy of nacre.
(A) SEM image of nacre. (B) Two distinct
Raman spectra of aragonite of different
crystal co-orientations (red and blue) with
characteristic peaks for the translational (153
rel. cm-1), wobbling (206 rel. cm-1) and CO-
stretching (1085 rel. cm-1) modes. (C) Overlaid
Raman and SEM image. The color code
corresponds to the colors of the spectra in (B).
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Tel. +49 (0) 731 140 700, Fax +49 (0) 731 140 70200
info@WITec.de, https://raman.oxinst.com

Automated polarization-resolved 0° 5° 10° 15° 20°

Raman imaging
In the next step, a series of Raman images
of the nacre with rotating polarization (5
degrees per step) was recorded while
maintaining a constant perpendicular 8 µm

25° 30° 35° 40° 45°

cross-orientation of the incident and
detection angles (Figure 13). Each of the
20 presented images in Figure 13 was
acquired from an area of 40 x 40 µm2
(160 x 160 Raman spectra, integration
time per spectrum 0.06 s). In the initial
state, the polarization of the incident 50° 55° 60° 65° 70°
light was parallel to the c-axis of the
aragonite crytal (Figure 13, 0 degrees).
The TrueComponent Analysis of the
data processing software was used to
evaluate the 160 x 160 Raman spectra
for 37 angular directions simultaneously
(a total of 947,200 Raman spectra) and 75° 80° 85° 90° 95°

identified three different components

(Figure 13). The red and blue areas
represent the co-oriented structures
similar to Figure 12C. Both show a strong
polarization dependence. Variations
in C-O stretching are apparent with
the green spectral component, which Figure 13: Polarization-dependent Raman imaging of nacre.
exhibits polarization effects opposite in Color-coded Raman images of nacre acquired at different polarization angles. Angle 0 degrees
angle from the blue and red components. reflects polarization perpendicularto the c-axis of the abalone nacre.

The underlying changes in the Raman

A B band intensities as a function of the
polarization angle are illustrated for
two sample positions for the entire
series of 37 angles in waterfall plots
(Figure 14). These representations of the
Raman spectra clearly show that lattice
Intensity [a.u.]

Intensity [a.u.]

vibrations of nacre are extinguished when

the polarization changes from 0 degrees
(perpendicular to the c-axis) to 45
degrees. At the same time, the intensity
of the C-O stretching band reaches its
maximum at this polarization angle.

Conclusion
Raman shift [rel. cm-1] Raman shift [rel. cm-1] Using the example of nacre, we
Figure 14: Polarization-dependent Raman spectral changes in nacre. demonstrated the immense potential
Waterfall plots of polarization-dependent Raman spectra in nacre. Individual spectra are polarization-resolved Raman ima-
given for 37 different polarization angles (5 degree steps). The spectral information is given for ging offers for obtaining a better
two individual positions representative for red (A) and blue (B) colored areas of Figure 13. The understanding of the crystal structures
coloration in the plots highlights the characteristic spectral features found in these areas. in biominerals. It is a fast, nondestructive
imaging technique, particularly suitable
for revealing information about crystal
orientation, unit cell distribution and
material composition.
TECHNICAL NOTE

Polarized White Light and Raman Microscopy on Biotite

Introduction Sample and underwent emplacement by melt
Since the invention of polarizers by William To differentiate geometrically hetero- of magmatic origin. Support for this
Nicol in 1829 and the development of the geneous biotite alignments, a sample hypothesis could be found in detecting
first polarizing microscope by Giovanni was chosen in which the relationship highly aligned biotite in the metamorphic
Battista Amici in 1830 [13,14], geosciences between two formation processes areas (pressure-induced solid reactions
and mineralogy have been revolutionized (metamorphic crystal alignment vs. and deformation), while seeing the
by the ability to analyze rock thin sections magmatic melt penetration) is not full degree of freedom for arbitrary
between a set of two polarizers with white obvious from crosscutting relationship. biotite crystallization (cavitation and
light illumination. Through the rotation Figure 15 shows this situation in a stitched crystallization from a liquid) in the
of the two polarizers relative to each white light image of a 40 x 20 mm thin magmatic areas.
other and to the orientation of individual section from the abandoned Griffins Find
mineral grains, the resulting optical Gold Mine, Western Australia. Two rock- Polarized white light microscopy
effects allow conclusions to be made forming generations could be assumed, investigation of biotite crystals
about materials‘s physical properties. as the metamorphic layering fabric of a With polarized white light microscopy,
Additional polarization-resolved Raman µm-sized mafic granulite (yellow region: optical properties of materials can be
spectroscopy measurements provide biotite-feldspar-pyroxene) is intersected visualized to aid in characterizing their
detailed insight on the crystallographic by a vein of mm-sized mineral grains composition and structure. The underlying
orientations in the rock. In this investigation with different mineralogical paragenesis effect – birefringence – is specific for
we combine both methods and (red region: quartz-feldspar-biotite- individual materials and is caused by the
semiquantitative approaches to identify pyroxene). However, it can also be seen anisotropism in the crystal lattice, splitting
two different formation stages of the that this vein propagates along the of the polarized light into two beam
mineral biotite based on its geometrical lineation and fabric of the metamorphic paths, and different propagation speeds
alignment in the sample. orientation. This raises the question of of the electromagnetic waves along
different crystal planes. This creates a
characteristic phase shift between both
waves, which can be represented as
interference colors.
Fig. 16 Here we show a 25 µm thin rock section
in different illumination setups: reflected
light (Figure 15A), transmitted light
(Figure 15B), transmitted light in parallel
i polarization (parallell-oriented polarizer
and analyzer, ||Pol, Figure 15C) and
cross polarization (orthogonal orientation
of polarizer and analyzer, xPol, Figure
ii 15D). While reflected and unpolarized
transmitted light images (Figures 15A and
15B) show the presence of opaque (ore
minerals) and transparent crystals (major
rock forming minerals), the coloring visible
Figure 15: Overview white light image of metamorphic rock with magmatic domain. in the ||Pol illuminated setting (Figure
15C) displays their intrinsic colors and,
Cross-polarization illumination showing a magmatic vein (red) crosscutting a metamorphic rock
along its pre-developed metamorphic fabric (yellow). White rectangles (i and ii) represent the
most importantly, their morphology and
selected areas for the following measurements; position of Figure 16 indicated with box. formative relationship. The interference
colors in the xPol configuration (Figure
whether this difference already existed 15D) indicate which minerals are present
(e.g. due to irregularities in the parent and reflects qualitative differences in
rock) or a possible melt penetration and the crystallographic orientation of their
recrystallization of the rocks occurred crystal lattices. In combination, the four
along the pre-developed lineation. images reveal mineral phase identities
A potential explanation for the sample’s as well as processes of rock formation
structure is that single crystals in by connecting optical properties to
a metamorphic rock were aligned crystallographic orientations and other
perpendicularly to the direction of crystal properties (i.e. crystal fabrics:
pressure over millions of years and habits, cleavages, twinning, inclusions,
then suddenly the rock was fractured exsolutions, deformations).
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A B Polarization-resolved Raman
spectroscopy of biotite
Raman spectroscopy was employed to
test our hypothesis on the origin of two
biotite generations in the rock. The first
approach was a statistical investigation
of two areas in the sample (respectively
Figure 15: white squares) that recorded
fully automated 360° polarizer series on
15 randomly selected biotite crystals in
both areas. A 50x/0.8NA objective and
a 532 nm excitation laser at 10 mW
were used. A series of single spectra was
C D acquired at an integration time of 0.5 s
per spectrum and 10 accumulations with
a polarizer angle step size of 5° through a
full rotation. Two selected Raman modes
of biotite and schematic representations
of their respective vibrations in the crystal
structure are given in Figure 17. Both, the
νTO4 stretching mode (T-O stretch of SiO4-
and AlO4-tetrahedron movements along
the c-axis, green vectors) and the δT6O6
ring modes (O3-T-O1 and T-O2-T bend
in the T6O6-ring adjacent to the cleavage
planes, blue vectors) were pre-evaluated
to be unaffected by birefringence effects
Figure 16: White light images of biotite mineral acquired using different illumination (Figure 17B) [15,16]. ▶
techniques.
Reflected (A) and transmitted (B) light image. Transmitted light image at parallel polarization
(||Pol) (C) and 90° crossed polarization setup (xPol) (D). Area section given in Figure 15.

A B
Intensity [a.u.]

Raman shift [rel. cm-1]

Figure 17: Structural formula, crystallographic models and polarization-resolved Raman spectra of biotite.
(A) Crystallographic models parallel to the b- and c-axis and structural formula of biotite. Coupling relationships are given for polarization
orientation parallel and orthogonal to the c-axis. (B) Resulting polarization-dependent Raman spectra which exaggerate T-O stretch parallel to
c-axis (SiO4- and AlO4-tetrahedron) and O3-T-O1 and T-O2-T bend orthogolalꞱ to c-axis (T6O6-ring).
TECHNICAL NOTE

T-O stretch O3-T-O1 bend + T-O2-T bend

i 90 90 ◀ For evaluation, polar orientation plots
120 60 120 60
were extracted displaying the Raman
α [°] α [°] band intensities of the νTO4 stretching
mode (green) and δδT6O6 ring modes (blue)
150 30 150 30

(Figure 18, 5 positions per area plotted).

180 0 180 0 In the metamorphic part (ii) a clear
orientation of the TO4-tetrahedrons‘ T-O
210 330 210 330
stretching mode along the 45° and 225°
polarizer rotation and thus parallel to
240 300 240 300 the former maximum pressure-induced
270 270
stress regime can be observed. The O3-
T-O1 and T-O2-T bending vibrations
ii 120
90
60 120
90
60 of the T6O6-rings, which correspond to
α [°] α [°] the anisotropy of the crystal lattice and
150 30 150 30 the preferred orientation of the layered
structure of the biotite parallel to the
180 0 180 0
(001) surface, are statistically regulated
along the 135° and 315° polarization.
In the magmatic part (i), however, the
210 330 210 330
crystal axes are irregularly oriented in all
spatial directions.
240 300 240 300
270 270

Figure 18: Polar plots of the biotite crystal orientations.

Measurement points and polar orientation plots of 5/15 polarizer series from the magmatic (i)
and metamorphic region (ii).

TrueComponent Analysis of palettes (inverse pole figures: red, blue and Conclusion
biotite in Raman images green), therefore correspond to spatial This study presented polarized white
In a second approach, high-resolution orientations in which the excitation laser light and polarization-resolved Raman
confocal Raman images were acquired is aligned perpendicularly to the biotite microscopy as effective methods for
from the magmatic (i) and metamorphic (100), (010) and (001) sectional planes investigating biotite crystals and their
(ii) areas and analyzed using the WITec and the laser polarization is aligned in crystallographic orientation. Using
TrueComponent Analysis data evaluation parallel with the crystallographic c-, a- semiquantitative approaches, the data
module. The images were recorded in a and b-axes, respectively. (Figure 19C). supported the hypothesis of different
2000 x 2000 µm area at 0.1 s integration The colors in the magmatically formed crystallization stages in the rock
time per spectrum using a 50x/0.8 NA area (i) are close to the extremes of formation. This clearly demonstrates
objective and a 532 nm excitation laser the color scale, indicating arbitrary the benefits of both techniques for
at 40 mW. In this analysis setup, only biotite orientations in this section that characterizing geoscience samples.
biotite spectra were included and three originate from random crystallization in
orientation endmembers (blue, green and a melt phase. In contrast, the image of
red; Figure 19A: selected endmember metamorphically regulated biotites (ii)
crystals, Figure 19D: endmember Raman shows a more uniform color spectrum,
spectra) were pre-defined as internal pointing to a similar orientation of all
standards for the crystallographic biotites in this area having been caused
orientations. Their allocation was based by high pressures applied over a long
on a detailed evaluation of the phase period of time.
identification and by the determination of In combination with the statistical Raman
their crystallographic axis orientation in spectroscopic approach, these results
xPol illumination. confirm a difference in the two parts of
As a result, the TrueComponent Analysis the thin section with highly ordered biotite
algorithm created intensity distribution crystals in the metamorphic and irregular
images (Figure 19B) in which the coloring orientations in the melt emplacement
of individual grains represents their areas, supporting the hypothesized
orientation match with the pre-defined manner of formation.
endmember orientations. The color
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Lise-Meitner-Str. 6, D-89081 Ulm, Germany
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info@WITec.de, https://raman.oxinst.com

A B C
i ii

(001) (001)

(010) (100) (010) (100)

D Figure 19: Raman imaging results made using TrueComponent

Analysis.
(A) Single crystals in parallel polarized white light image
Intensity [a.u.]

selected for endmember definition (red, blue and green). (B)

pol || c TrueComponent Analysis results expressed as false color images
matching the (100), (010) and (001) planes of biotite. (C) Crystal
pol || a models and geometric alignment of the endmembers towards
the laser polarization vector. (D) Corresponding endmember
pol || b Raman spectra extracted from (A).
Raman shift [rel. cm-1]

References
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correlativeImaging.pdf
[2] Xu, B., Mao, N., Zhao, Y., Tong, L., & Zhang, J. (2021). Polarized Raman spectroscopy for determining crystallographic orientation of low-
dimensional materials. The Journal of Physical Chemistry Letters, 12(31), 7442-7452.
[3] Allen, V., Kalivas, J. H., & Rodriguez, R. G. (1999). Post-consumer plastic identification using Raman spectroscopy. Applied spectroscopy, 53(6), 672-
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Polaritons in a van Der Waals Crystal. Advanced Materials, 33(2), 2005777. DOI: 10.1002/adma.202005777.
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[8] Zheng, B., Wang, Z., Chen, Y., Zhang, W., & Li, X. (2018). Centimeter-sized 2D α-MoO3 single crystal: growth, Raman anisotropy, and optoelectronic
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[9] Seguin, L., Figlarz, M., Cavagnat, R., & Lassègues, J. C. (1995). Infrared and Raman spectra of MoO3 molybdenum trioxides and MoO3· xH2O
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8539(94)00247-9.
[10] Loh, H. C., Divoux, T., Gludovatz, B., Gilbert, P. U., Ritchie, R. O., Ulm, F. J., & Masic, A. (2020). Nacre toughening due to cooperative plastic
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[12] Farfan, G. A., Zhou, C., Valley, J. W., & Orland, I. J. (2021). Coupling mineralogy and oxygen isotopes to seasonal environmental shifts recorded in
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