რ

Life, Earth andPlanetary Sciences

Ⅲ-5 Life, Earth and Planetary Sciences

BL3U

X-ray Absorption Spectra of Lipid Bilayer Membranes in Aqueous Solutions
R. Tero 1, Yu Kinjo1 and M. Nagasaka2
1

Toyohashi University of Technology, Toyohashi, 441-8580, Japan
2
Institute for Molecular Science, Okazaki 444-8585, JapanThe lipid bilayer is a self-assembled structure ofamphiphilic lipid molecules. It is the fundamental
structure of biomembranes such as cell membranes,
which are the fields for the transportation of materials,
information, and energy into and out of cells. All these
physiological reactions proceed in the presence ofelectrolytes. Ions in the aqueous solution significantly
influence to physical properties and structures of lipid
bilayers. Phosphatidylcholine (PC) is the most abundant
lipid of eukaryotic cell membranes. Cations bind to thephosphate and carbonyl groups of PC. However,aﬃnity of cations to PC and eﬀects of cations tomolecular orientation are still controversy especially inthe fields of theoretical simulations. We aim todetermine the binding aﬃnity of cations to lipids inaqueous solutions experimentally, by means of X-ray
absorption spectroscopy (XAS) [1, 2].
Supported lipid bilayers (SLBs) of dioleoyl-PC(DOPC) were formed on the Si3N4 membranes of the
XAS flow cell [1] by the vesicle fusion method in a
buﬀer solution (NaCl 100 mM, HEPES 25 mM/ pH 7.4
NaOH). The Na+ concentration ([Na+]) was varied byexchanging the buﬀer solution in the flow cell in in therange of [Na+] = 2.1 – 510.4 mM. The O-K edge XAS
spectra of SLB were obtained at the energy range of
527 – 535 eV at the Xray incident angle of 35º. The
XAS spectrum of the Si3N4 membrane without SLB
was subtracted from that with SLB.
The O-K-edge spectrum of DOPC appeared at 531 533 eV (Fig. 1). It is consisted of 1s→π* transitions ofdouble-bond oxygens in the phosphate and carbonylgroups on the PC headgroup: two components attributed
to the P=O in the former, and one component attributed
to the latter [3]. We measured XAS spectra at the X-ray
incident angle (T) of 35º to obtain the intensity of eachcomponent independently of the molecular orientation.
We obtained a specific dependence of the P=O peak at
the lower energy (P=O_low) on [Na+]: its position was
identical at [Na+] = 2.1 – 10.4 mM, shifted to higher
energy by 0.4 eV above [Na+] = 10.4 mM, and little
independent of [Na+] above 110.4 mM. The results
show the Na+ coordination to the phosphate group of
PC and its dependence of [Na+]. The inner-shell
quantum chemical calculation [4] of O K-edge spectrumof P=O indicated that coordination of Na+ to thephosphate group causes higher-energy shift of P=O
peaks. The dependence of the P=O peak intensity as
well as the P=O intensity on [Na+] is investigated indetail. The eﬀects of the cation species on the XAS
components will be investigated.

Fig. 1. O K-edge XAS spectra of DOPC-SLB at [Na+] =2.1 – 510.4 mM obtained at the X-ray incident angle of
35°. Black dotted curves represent raw data. Dashedcurves and red solid curves show deconvoluted
components and their summation, respectively.
[1] M. Nagasaka, H. Yuzawa, T. Horigome, and N.
Kosugi, J. Electron Spectros. Relat. Phenomena 224(2018) 93.
[2] R. Tero, Y. Kinjo and M. Nagasaka, UVSOR
Activity Report 49 (2021) 151.
[3] R. Tero, W.-Z. Goh and M. Nagasaka, UVSOR
Activity Report 48 (2020) 129.
[4] M. Nagasaka, J. Chem. Phys. 158 (2023) 024501.

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BL1U

Study on Chirality Emergence of Amino Acid Molecules Induced by
Circularly Polarized Lyman-α Light Irradiation
M. Kobayashi1, J. Takahashi2, H. Ota3, G. Fujimori2, K. Kobayashi2, K. Matsuo4, Y. Taira3,
M. Katoh3,4, H. Nakamura1 and Y. Kebukawa2
1

National Institute for Fusion Science, 322-6 Oroshi-cho, Toki 509-5292, Japan
Yokohama National University, 79-5 Tokiwadai, Hodogaya-ku,Yokohama 240-8501, Japan
3
UVSOR Synchrotron Facility, Institute for Molecular Science, Myodaiji,Okazaki 444-8585, Japan
4
Hiroshima Synchrotron Radiation Center, Hiroshima University, 2-313 Kagamiyama,
Higashi-Hiroshima 739-0046, Japan2

The origin of homochirality in terrestrial biomolecules(L-amino acid and D-sugar dominant) remains one of
the most mysterious problems in the research for theorigins of life. One of the most attractive hypotheses in
the context of astrobiology is “Cosmic Scenario” [1, 2];Asymmetric reactions of prebiotic molecules oninterstellar dust surfaces in molecular cloud
circumstances were introduced by polarized quantum
radiation sources in space, that is “chiral radiations”.
Among the polarized quantum radiation sources,
circularly polarized light (CPL) in the spaceenvironment is thought to be one of the most likely
causes of the enantiomeric excesses of terrestrial
bioorganic molecules. In this study, we focused on ahydrogen Lyman-α wavelength of 121.6 nm, wherestrong emission lines are observed in star-forming
regions.
Alanine is selected as a chiral prebiotic molecules in
the interstellar space. We formed thin solid film samplesof racemic mixture of alanine (DL-alanine) on quartz
substrates from crystal powders of DL-alanine by using
a thermal-crucible vacuum-evaporation system at
HiSOR in Hiroshima university. A schematic of theirradiation system of BL1U is shown in Fig.1. The
121.6 nm wavelength radiation from the undulator isreflected by a gold-coated mirror (M0 focusing mirror)
located in the mirror chamber directly beam upstreamof the irradiation chamber and then enters the irradiationchamber. On the beam entrance side of the vacuum
sample chamber, a gate valve with an MgF2 vacuum
sealing window (0.5 mm in thickness) was mounted.
The use of gold-coated mirror reflections has made it
possible to suppress high-energy higher order lightfrom the undulator source, expecting to reduce thetransmittance loss of the MgF2 window due to highenergy radiation induced defects. The sample substrate
was set in the sample holder in the irradiation chamber,in which magnetic and electric fields can be applied toperpendicularly to the sample surface. The total photon
beam intensity irradiated on the sample was monitoredwith photoelectron current of a silicon photodiodesettled at the beam downstream side of the sample
holder.
CD spectra of the CPL irradiated films were measured
using the SR-CD beam line BL-12 of HiSOR to clarify

the optical activity emergence by CPL irradiation.
Figure 2(a) shows spectra of DL-alanine films irradiatedfor 30 min with L- or R-CPL at 121.6 nm in wavelength.
We clearly observe optical activity induced by the CPL
irradiation, which shows opposite sign in the CD
between right CPL and left CPL. In addition, we havealso examined the additional eﬀect of applying amagnetic field to the sample to investigate the eﬀect ofthe magnetic field in interstellar space (Fig. 2(b)). The
application of the magnetic field seems to alter the CD
spectra, suggesting a certain impact of symmetry
breaking of the optical activity caused by the CPL
irradiation. Detailed analysis of CD spectra is inprogress supported by quantum chemical calculations.

Fig. 1. A schematic of the CPL irradiation system at
BL1U.

Fig. 2. CD spectra of alanine sample after irradiation of(a) right (red) and left (black)-CPL, and (b) right CPL
with magnetic field application (blue:+0.7T, green:0.7T).
[1] W. A. Bonner, Orig. Life Evol. Biosph. 21 (1991)
407.
[2] J. Takahashi and K. Kobayashi, Symmetry 11(2019) 919.

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Ⅲ-5 Life, Earth and Planetary Sciences

BL3U

Attempts at Structural Analysis of Biological Macromolecules Using
Resonant Soft X-ray Scattering
Y. Sugimoto1 and H. Iwayama1,2
1
UVSOR Synchrotron Facility, Institute for Molecular Science, Okazaki 444-8585, Japan
School of Physical Sciences, The Graduate University for Advanced Studies (SOKENDAI),
Okazaki 444-8585, Japan

2

Resonant soft X-ray scattering (RSoXS) is a powerful
technique to probe the nano-to-mesoscopic structure ofmolecules. Since this method observes scattered waves
due to resonance processes, RSoXS can reveal
structures with enhanced or tunable scattering contrast.
In particular, since soft X-ray regions include K-edgeenergies of light elements such as carbon, nitrogen, andoxygen, RSoXS is a suitable tool for investigating soft
matters. In UVSOR, the development of the RSoXS
instrument has begun since 2019, and we havesuccessfully measured the mesoscopic structure ofliquid crystal molecules [1].
Biomacromolecules, such as proteins, are also mainlycomposed of light-element atoms. Studying
biomacromolecule structures is crucial for elucidating
biological functions and developing therapeutic drugs.
Conventional electron microscopy (EM) and smallangle X-ray scattering (SAXS) are well-establishedmethods for investigating the structure of biologicalmacromolecules. The RSoXS method is expected tocomplement conventional methods due to its features,such as the selectivity of elements and functional grouporientations. In this work, we performed RSoXS
measurements for protein samples.
Our RSoXS measurements were performed at
UVSOR BL3U. Our sample is bovine milk casein as
the typical protein sample. Casein form micelles withcalcium phosphate dispersed inside [2]. We prepared
the bovine milk samples in the following step. First, wedissolved the bovine milk powder sample in water, thendropped samples onto a Si3N4 membrane and driedthem naturally. We set the samples in the vacuum
chamber and irradiated them with soft X-rays. We
measured the scattering intensity with a soft X-ray
CCD camera (Andor BN940P). The photon energies
were set to 270, 285, 348, and 350 eV. The measured
Q-range was 0.035-0.35 nm-1 at 270 eV and 0.055-0.45
nm-1 at 350 eV. The Guinier plot of the scattering
intensity yields the radius of gyration of ~55 nm,

measured at 270 eV below the carbon K-edge
absorption. This value is close to the structure of casein
micelles obtained in previous studies. The scattering
intensity measured at 350 eV, near the calcium L
absorption edge, showed a diﬀerent scattering intensity
than that at 348 eV (Fig. 1). The resonance-enhanced
structure of the spectrum suggests that RSoXS gives
insight into a more detailed structure of calcium
phosphate in casein micelles, compared to conventional
SAXS.
On the other hand, some structural changes inbiomolecules may come from drying process of thesample preparations. In addition, our equipment isdiﬃcult to measure small scattering-angle regions dueto the direct beam. We will introduce a new sample cellfor wet samples and improve our RSoXS spectrometerfor small scattering angle measurements.

Fig. 1. Q-spectra for casein at the calcium L absorptionedge.
[1] Y. Takanishi et al., RSC Adv. 12 (2022) 29346.
[2] H. Takagi et al., Food Chem. 393 (2022) 133389.

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BL3U

Water-Ion Interactions in Natural Lake Brines
1

Y. Hao , N. Fauré2, Y. Qiu1, M. Nagasaka3, E. Thomson2, S. Wang1 and X. Kong2
1

Department of Environmental Sciences, College of Urban and Environmental Sciences,
Northwest University, Xi’an, China
2
Department of Chemistry and Molecular Biology, Atmospheric Science, University of Gothenburg,
SE-412 96 Gothenburg, Sweden
3
Institute for Molecular Science, Okazaki 444-8585, Japan

Under climate change and enhanced water use
demands, arid regions are undergoing fastertransformations compared to humid areas due to theirgreater susceptibility to changes in precipitation. As aconsequence, the number and area of saline lakes arebeing impacted. Depending on the local circumstances,some saline lakes are being desiccated and transforming
to playa covered by evaporite salts, while others are
forming and expanding, such as the lakes in the
Qinghai-Tibet Plateau [1]. The increases volume andsurface area of saline lakes are expected to have impacts
on local environment and climate.
In this beamtime, we carried out Near Edge X-ray
Absorption Fine Structure (NEXAFS) measurementson brines from various saline lakes in the Qaidam
Basin, which is situated in the Qinghai-Tibet Plateau.
The sampling sites include eight saline lakes: Mang’ai
Feicui Lake (MA), Dalangtan Playa (DLT), West
Taijinar Lake (XT), East Taijinar Lake (DT), Qarhan
Lake (QH), Dezongmahai Lake (DZMH), Dachaidan
Lake (DCD), and Xiaochaidan Lake (XCD) (Figure 1).
The brine samples were taken 2 – 5 cm below the lake
surface, and then were stored in polyethylene bottles,
sealed with Parafilm membranes, and stored at 4 °C.
The NEXAFS measurements were performed at thesoft X-ray undulator beamline BL3U at UVSOR-Ⅲ
Synchrotron. In the liquid flow cell, a liquid layer issandwiched between two 100 nm thick Si3N4
membranes with the windows size of 2 × 2 mm2. Teflon
spacers with the thickness of 100 μm are set between
the support plates of the Si3N4 membranes. Liquidsamples are exchangeable in situ by using a tubing
pump. The temperature of all the liquid samples were
controlled at approx. 25 °C. The Si3N4 membrane
window was set to 200 × 200 μm2 to allow for suitable
photon fluxes.
In Figure 2, the O K-edge NEXAFS results arepresented, with 8 brines displayed in two panels forbetter visibility. The pure water case is also included inboth panels for comparison purposes. The spectra
exhibit three features: pre-edge (~535 eV), main-edge(~537 eV), and post-edge (~540 eV), with the spectrum
lines normalized to the maximum at the main-edge.
The post-edge intensities of the brines are lower thanthose of pure water, which is consistent with existing
literature [2]. The pre-edges have been zoomed in anddisplayed in the insets. Apart from the DZMH brine,
the other brines exhibit reduced and shifted pre-edge

peaks, which will be analyzed in more detail. The
DZMH brine displays a notably high pre-edge, likely
resulting from contamination.

Fig. 1. Eight saline lakes, as marked by red circles,
were sampled in the Qaidam Basin.

Fig. 2. O K-edge NEXAFS spectra of (a) pure waterand brines from MA, DLT, DCD, and QH lakes; (b)pure water and brines from DT, DZMH, XT, and XCD
lakes. The temperatures are around 25°C. The insets
zoom in the pre-edge regions.
[1] Zhang, G. et al., Earth Sci. Rev. 208 (2020) 103269.
[2] Nagasaka, et al., J. Phys. Chem. B, 121 (2017)10957-10964.

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Ⅲ-5 Life, Earth and Planetary Sciences

BL4U

Drug Administration Concepts Involving Redox Nanocarriers Aﬀect theSpatial Distribution of Topically Applied Formulations in Human Skin
G. Germer1, T. Ohigashi2, H. Yuzawa2, K. Rajes3, R. Haag3, F. Rancan4, A. Vogt4 and E. Rühl1
2

1
Physical Chemistry, Freie Universität Berlin, Arnimallee 22, 14195 Berlin, Germany
UVSOR Synchrotron Facility, Institute for Molecular Science, Okazaki 444-8585, Japan
3
Organic Chemistry, Freie Universität Berlin, Arnimallee 22, 14195 Berlin, Germany
4
Charité Universitätsmedizin, 10117 Berlin, Germany

Diﬀerent drug administration concepts involving
redox-sensitive polymer nanocarriers topically applied
to human skin ex vivo have been investigated byscanning transmission X-ray microscopy (STXM). The
goal of this research is to understand how single andrepeated use of drug formulations containing
nanocarriers aﬀects the drug and nanocarrier
distribution in the top skin layers of inflamed skin.
Especially, the repeated use of drug formulations isfrequently used in therapy. STXM probes sensitively
and label-free the distribution of topically applied
formulations [1]. The results are of importance tounderstand the transport and penetration routes ofactives. Polymer nanocarriers are used for transporting
drugs that are barely penetrating the skin barriers [2, 3].
The present studies compare diﬀerent topical drugdelivery concepts of a single dose and repeated drug
administrations, reaching up to five days, respectively.
The skin barrier was initially modified for all samples
by topically applying the oxidizer di-benzoyl peroxide(DBPO) for 16 h prior to topical application of
rapamycin-loaded oxidation-sensitive nanocarriers(osCMS [2]). The osCMS samples (5 mg/mL, ≤2%drug loading) were formulated in HEC gel. 40 μL/cm2of this formulation were applied once for up to 1000
min to the skin samples ex vivo. In case of repeatedtreatments, the skin was four times treated every 24 hwith 40 μL/cm2 of a 1% rapamycin-osCMS formulation
with a final drug concentration of 8 μg/cm2. The STXM
experiments were performed at the BL4U beamline of
UVSOR III at the O 1s regime (520 – 565 eV). Key isthe acquisition of stacks of images by concentrating on
the top skin layers, i.e., the stratum corneum (SC) andthe viable epidermis (VE). Fig. 1 shows selected results,
where a comparison is made between the single
treatment (Fig. 1(a)) and multiple treatments (Fig.
1(b)). At the top of each Figure an X-ray micrographrecorded at 532 eV is shown, indicating the skinstructure of the vertical cuts of EPON-fixed skin, where
the skin surface is located at the right hand side. Each
Figure also contains the local concentrations of
rapamycin ① and osCMS ②, derived from theexperimental results and subsequent singular value
decomposition, similar to recent work [1]. Single
treatment shows primarily the species of interest in thestratum corneum (Fig. 1(a)), where the nanocarriers aremostly found in narrow spatial distribution in the lipids
separating the corneocytes, whereas the drug is foundin the corneocytes. Multiple treatments (Fig. 1(b))
show a diﬀerent distribution of both species: Most of
the drug has been transported in deeper skin layers,
only some of the rapamycin remains in corneocytes.
Nanocarriers are primarily found in the viableepidermis, and there they are localized near the cell
nuclei. This deep penetration of osCMS into the viable
parts of skin is likely the result of a damaged tightjunction barrier in the stratum granulosum due to initial
treatment by DBPO, as nanocarriers were not observed
before in the viable parts of intact skin after single
treatment [3]. These diﬀerences in drug and nanocarrier
distributions provide evidence that in inflamed skin the
drug is eﬃciently transported via nanocarriers to thesites of action, where it is needed for therapeutic
success, considering that the drug dissolved in ethanol
is barely taken up by intact skin [1].

Fig. 1. Results from STXM on diﬀerent drug delivery
concepts of human skin ex vivo initially treated by
DBPO: (a) single drug application (1000 min); (b)repeated use of drug administration (5 d). Top: X-ray
micrograph recorded at 532 eV, below: local
concentrations of ① rapamycin and ② osCMS. Scalebar: 5 μm.
[1] G. Germer et al., ACS Omega 6 (2021) 12213.
[2] K. Rajes et al., Pharmaceutics 13 (2021) 37; K.
Rajes et al., Biomater. Sci. Engin. 7 (2021) 2458.
[3] K. Yamamoto et al., J. Control. Release 242 (2016)64.

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BL4U

Absorption- Image and Spectra of Apoptosis Nuclei in P-L Edge
T. Ejima1 and S. Tone22

1
SRIS, Tohoku University, Sendai 980-8577, Japan
Sch. of Sci & Technol., Tokyo Denki University, Hatoyama 350-0394, Japan

Apoptosis is programmed death of cells that is an
important physiological function for the recycling ofcellular components and characterized by chromatincondensation in the cell nucleus. Detailed studies of thechromatin condensation process in isolated cell nuclei
have shown that this condensation process occurs inthree stages from the normal state of the cell nucleus(Stage 0) to ring condensation (Stage 1), necklace
condensation (Stage 2), and cell nuclear collapse/
disassembly (Stage 3) [1]. It has been suggested thatamong the transitions from Stage 0 to 3, Stage 1 to 2 iscaused by cleavage of phosphodiester bonds in DNA,
while protein kinases such as MST1 may be thecohesive factor in the transition from Stage 2 to 3 [1].
To investigate the change from Stage 0 to Stage 1,
infrared absorption experiments were performed,
focusing especially on the change in the vibrational
structure of PO2−, which is sensitive to the helicalstructure of DNA. As a result, the helical structure,
which was predominantly B-DNA in Stage 0, decreased
as the step progressed, and changed to A-DNA or
Z-DNA. This result suggests that the helical structureof DNA changes from Stage 0 to 1, DNA is cleaved inStage 1 to 2, and phosphate groups are transferred from
DNA in Stage 2 to 3. In this study, we focused on thischange in the phosphate group of DNA and thought that
we could capture the changes in DNA at each stage bymeasuring the P-L2,3 absorption edge structure of thecell nucleus.
The samples used in the experiments were obtained
according to the method reported by Tone et al [1]. The
isolated nuclei of each stage obtained were embedded
and sectioned into approximately 100 nm-thick section
samples using an ultramicrotome. The sectioned
samples were imaged under an optical microscope andcell nuclei showing representative shapes of each stage
were selected in the images. Isolated nuclei of Stage 0
and Stage 2 were selected for measurement due to time
limitations, and STXM measurements of the C-K and
P-L absorption edges were performed for each. The
obtained STXM images were analyzed by PCA andreconstructed after removing noise components fromthe PCA component images, and cluster analysis was
performed to the reconstructed images [2]. Figure
shows the STXM image of a Stage 2 isolated nucleus at
the P-L absorption edge and the obtained absorption
spectrum. The spectra show two peak structures, P-L2
and L3, split by spin-orbit interaction. We believe thatthe spectral shape of each peak is similar to that of PO4.
In combination with the image, it is considered that
DNA was degraded and transformed into PO4 clusters
as apoptosis progressed. This interpretation is consistent
with the result of IR absorption results.

Fig. 1. X-ray absorption image, (a), and X-ray
absorption spectrum, (b), of apoptosis nucleus in stage2
at P-L absorption edge. Bar is 2 μm in length.
References:[1] S. Tone et al., Exp. Cell Res. 313 (2007) 3635.
[2] M. Lerotic et al., JESRP 144-147 (2005) 1137.

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Ⅲ-5 Life, Earth and Planetary Sciences

BL4U

Embracing Natural and Anthropogenic Aerosol Particle Complexity
P. A. Alpert1, L. Li2, L. F. Escusa Dos Santos2, F. Mahrt1, M. Ammann1, L. Wu3, W. Hu3,
P. Fu3, M. Hallquist2, E. Thomson2 and X. Kong2
1

Laboratory for Environmental Chemistry, Paul Scherrer Institute, Villigen PSI, 5232, Switzerland
2
Atmospheric Science Research Group, Department of Chemistry and Molecular Biology,
University of Gothenburg, 41296 Gothenburg, Sweden
3
School of Earth Systems Science, Tianjin University, 300072 Tianjin, China

A considerable fraction of the global population lives
in coastal urban centers. People in these regions are
exposed to some of the most complex aerosol particles
containing mixtures of natural and anthropogenic
compounds. The chemical composition and morphologyof these particles are key for their environmental
impacts, but remain poorly understood. We haveinvestigated particles taken from ambient air that was
influenced by the ocean and mixed ocean-anthropogenic
sources to better understand their cloud-forming
potential, chemical reactivity and light-scattering
properties.
Sea spray aerosol particles contain sea-salts andorganic matter [1]. When mixed with anthropogenic
air, these particles can uniquely undergo aciddisplacement reactions forming organic salts andproducing inorganic acids, which evaporate from theparticles [2]. Mixed anthropogenic and natural aerosol
particles aerosolized from saline lakes have previously
been shown to act as cloud nuclei [3]. Additionally,
anthropogenic emissions from ships are known to havea variable ability to nucleate clouds depending on thetype of fuel used and its sulfur content [4].
To better understand such complex processes wecollected ambient particles from contrasting sources, i)
the Indian Ocean with minimal anthropogenic
influence, ii) the Indian Ocean with significant
anthropogenic influence and iii) Sanya Port, a coastal
city in China. At the Ultraviolet Synchrotron Orbital
Radiation Facility, we probed individual particles at the
carbon, oxygen and nitrogen K-edges, and the sulfur,
chlorine and calcium L-edges using scanningtransmission X-ray microscopy coupled to near-edge
X-ray absorption fine structure spectroscopy (STXM/
NEXAFS). We quantified how inorganics and organics
were mixed in individual particles and determined theorganic volume fraction (OVF) [5]. The OVF was
calculated from high spatial resolution images at 280
eV, 285.4 eV, 288.7 eV and 320 eV with a pixel size of
0.05 × 0.05 μm2.
Figure 1 shows optical density measured at 280 eV
and OVF for the same field of view of Indian Ocean
particles without anthropogenic influence. The opticaldensity image shows X-ray absorption primarily due toelements that are not carbon, such as Na and Cl, as wellas other sea salts and inorganic matter. Particle
morphology is largely cubic with submicrometer sizedneedle-like structures, as expected for sea spray aerosol
particles. Based on our STXM/NEXAFS measurements,
these needles were confirmed to be dominated bycalcium sulfate. The OVF image reveals a considerable
fraction of organic material close to salt cores. Carbon
K-edge spectra were dominated by carboxyl functionalgroups, similar to sea spray aerosol particles found overvarious ocean environments [1, 2, 5].
These data suggest that these particles will likely behighly hygroscopic and act as cloud condensation and
ice nuclei due to their high salt content and similar
carbon spectra as other ice nucleating particles [1].
Additionally, morphology and composition determinedfrom STXM/NEXAFS are similar to laboratorygenerated ice nucleating sea spray particles, implying arole for atmospheric ice particle production [1]. The
highest OVF appears close to the salt at values around
0.6 and decreases with distance away from the crystal.
This implies a small degree of aging, i.e., that organic
salts dominate likely due to acid displacement reaction
rather than pure organic coatings. Further work is
ongoing to identify diﬀerences in particles from various
sources and their importance for cloud microphysics
and aerosol chemistry.

Fig. 1. Optical density at 280 eV indicating X-ray
absorption primarily due to compounds other thancarbon. Organic volume fraction of the same particles
calculated for particles collected over the Indian Ocean.
Scale bar is 2 μm.
[1]P. A. Alpert et al., Sci. Adv. 8 (2022) eabq6842.
[2]A. Laskin et al., J. Geophys. Res.-Atmos. 117 (2012)
D15302.
[3]J. Li et al., Atmosphere 12 (2021) 1203.
[4]L. F. E. d. Santos et al., Environ. Sci.: Atmos. 3(2023) 182.
[5]M. Fraund et al., Atmos. Meas. Tech. 12 (2019)1619.

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STXM in Characterizing Lysosomal Storage Materials in a Mouse Model
T. Mansikkala1,2 , T. Ohigashi3, H. Yuzawa3, I. Miinalainen2, S. M. Kangas4, M. Salo4,5,
N. Kosugi3, J. Uusimaa4, M. Huttula1, R. Hinttala 4,5 and M. Patanen1, 2
1Nano and Molecular Systems Research Unit, PO Box 3000, 90014 University of Oulu, Finland
2
Biocenter Oulu, PO Box 5000, 90014 University of Oulu, Finland
3
UVSOR Synchrotron Facility, Institute for Molecular Science, Okazaki 444-8585, Japan
4
Research Unit of Clinical Medicine and Medical Research Center, Oulu University Hospitaland University of Oulu, 90014 Oulu, Finland
5
Transgenic and Tissue Phenotyping Core Facility, University of Oulu, Finland

In this work, we used scanning transmission X-ray
microscopy (STXM) technique at BL4U beamline to
study lysosomal storage disease (LSD) mouse model
liver and kidney tissues. LSDs are a large family of
diseases where e.g. enzymatic dysfunction leads toaccumulation of macromolecules in the tissues. This
study was part of a project to better understand them.
STXM hold promise to be a good tool to study LSDs if
the natural chemical contrast of accumulating material
and rest of the tissue is high enough. In our previous
beamtimes at UVSOR both human and mouse model
samples have been studied (Patanen 2018, Mansikkala2020). This time our aim was to diﬀerentiate the storage
material within the lysosomes. Liver and kidney tissue
samples were used. They both had clear ultrastructural
diﬀerences compared control samples in form of large
number of vacuoles, presumed to be lysosomes fitting
the description of the LSDs. Our preliminary studies
showed a distinct spectrum from lysosomes comparedto the rest of the tissue. This study was to verify thefinding in larger number of samples.
The tissue samples were fixed in 4% paraformaldehyde
and 2.5% glutaraldehyde in 0.1 M phosphate-buﬀeredsaline and then embedded in TTE:MMHA. The 150 nm
thick sections were imaged with TEM to find areas withlysosomes on the samples. The same area was thenmeasured with STXM from the subsequent thinsections.
We were able to find and image lysosomes in bothtissue types, an example shown in Fig. 1. When
analyzing the samples with aXis2000, we noticed thatthe spectrum from the lysosomes was very similar tothat of the TTE:MMHA resin (Fig. 2). To evaluate theresult, we subtracted the measured pure resin spectrum
from the measured lysosome spectrum shown in Fig. 3(green). There is only a small peak around 289 eV
diﬀerentiates the lysosome spectrum from a resin
spectrum. However, the spectrum from the extracellularspace (blue) is almost identical with just intensity
diﬀerences. The spectrum of cytosol diﬀers clearly,
indicating that the contents of the lysosomes are mostly
TTE:MMHA resin. While there is a small diﬀerence
between the resin and lysosomes and extra cellularspace, the spectra are overwhelmed by the resin signal,
and we were not able to diﬀerentiate the accumulationmaterial spectrum in the lysosomes. Our hypothesis isthat the sample preparation has substituted theaccumulation material in lysosomes with resin insteadof preserving it.

p

Fig. 1. STXM image of disease model mouse kidneywith visible ultrastructural changes. Lysosomes aremarked with 1, cytosol with 2 and extracellular spacewith 3. Scale bar is 5 μm.

Fig. 2. Comparison of spectra from lysosome (blue)
and pure resin (red).

Fig. 3. Spectra from lysosome (green), the extracellularspace (blue) and cytosol (red).
[1] T. Mansikkala et al., UVSOR Activity report 2019(2020) 155.
[2] M Patanen et al., UVSOR Activity report 2018(2019) 166.

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Ⅲ-5 Life, Earth and Planetary Sciences

BL4U

Determination of Spectral Changes in NEXAFS of Biomolecules Caused by
Radiation Eﬀects for Molecular Mapping of Biological Specimens
A. Ito1, K. Shinohara2, A. Matsuura3, S. Toné4, K. Tohya5, H. Yuzawa6 and T. Ohigashi6, 7
1
School of Engineering, Tokai University, Hiratsuka 259-1292, Japan
Graduate School of Health Sciences, Fujita Health University, Toyoake, 470-1192, Japan
3
Research Integrity Oﬃce, Fujita Health University, Toyoake, 470-1192, Japan
4
School of Science and Engineering, Tokyo Denki University, Hatoyama, 350-0394, Japan
5
Kansai University of Health Sciences, Kumatoricho, 590-0482, Japan
6
UVSOR Synchrotron Facility, Institute for Molecular Science, Okazaki 444-8585, Japan
7
School of Physical Sciences, The Graduate University for Advanced Studies, Okazaki 444-8585, Japan2

1.4(Pm2/pg)

Mass absorption coefficient

(a)

(b)

1.2
1
0.8
0.6
0.4
0.2
0

281 eV
0

0.5

1

1.5

Absorbed dose (MGy)(Pm2/pg)

Mass absorption coefficient

1.4

(c)

1.2
1
0.8
0.6
0.4
0.2
0

393 eV
0

20

40

60

80

100

Absorbed dose (MGy)

120

1.4

(Pm2/pg)

Mass absorption coefficient

We have been working on a quantitative mapping of
nucleic acids (DNA and RNA) and proteins (histone
and bovine serum albumin abbreviated as BSA) in
chromosomes, cultured mammalian cells, and isolated
apoptotic nuclei using combined NEXAFS spectra at
the C, N and O K-edges [1-3]. The molecular mappingis based on the NEXAFS profiles of constituent
biomolecules obtained independently. However,increased exposure dose required for X-ray imaging
will modify NEXAFS profiles of biomolecules asreported previously [4]. For the precise molecularmapping of biological specimens, spectra measured at
the same exposure as that used in the specimen imaging
are required. Furthermore, the knowledge of the
exposure dependence of NEXAFS profiles would
improve the reliability of molecular mapping of
biological specimens whose images are taken at any
exposure time.
In our previous report, we detected a slight change of
NEXAFS profiles of DNA at C, N and O K-edges,
depending on the exposure time [4]. In the present
study, we extended our analysis to the absorbed dose
dependence of the mass absorption coeﬃcient for all
the energy points of NEXAFS, where absorbed dose iscalculated from the exposure.
We prepared dry thin films of DNA, RNA, histone,
BSA, actin, actin monomer and nucleosome on SiN
membrane. Each one micrometer square area in thespecimens was scanned with 10 x 10 pixels at theenergy of C, N and O K-edge regions mainly in thisorder with a dwell time of 6, 12 and 20 msec in addition
to 2 msec for DNA. For each dwell time the scannedarea was shifted to the diﬀerent area to avoid additionalradiation eﬀects to molecules.
Examples of the absorbed dose dependence of mass
absorption coeﬃcient are shown in Figure 1 at thelowest energies in the NEXAFS measurements of DNA
at the C, N and O K-edges. All data in the repeatedexperiments for several times were compiled in the
figures. The linear regression lines showed a slightdecrease with increasing absorbed dose, which meansthat radiation aﬀects the spectra of molecules.
Estimation of NEXAFS profiles at a given absorbed
dose is in progress.

1.2
1
0.8
0.6
0.4

524 eV

0.2
0

0

40

80

Absorbed dose (MGy)

120

160

Fig. 1. Eﬀects of soft X-rays on mass absorption
coeﬃcients in DNA at (a) C K, (b) N K, (c) O K- edges.
The solid lines show linear regression.
[1] K. Shinohara et al., Ultramicrosc. 194 (2018) 1.
[2] K. Shinohara et al., J. X-Ray Sci. Technol. 26(2018) 877.
[3] K. Shinohara et al., Cells 8 (2019) 8.
[4] A. Ito et al., UVSOR Activity Report 49 (2021) 152.

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BL4U

Attempt to Constrain the Initial Condition of Aqueous Alteration
on Cometary Organic Matter
H. Yabuta1, N. Nishii1, T. Araki2 and H. Yuzawa2
1

Department of Earth and Planetary Systems Science, Hiroshima University, Kagamiyama 1-3-1,
Hiroshima 739-8526 Japan
2
UVSOR Synchrotron Facility, Institute for Molecular Science, Okazaki 444-8585, Japan

This study aims to understand the chemical evolutionof organic matter in the early stage of aqueous alterationof icy small bodies, through the freeze-thaw experimentof analogues of cometary organic matter.

(UCAMM) [3], supporting the formation of the
UCAMM with a very small amount of fluid on acometary body.

As a starting material, HCN polymer was synthesized
by heating formamide (0.3 ml) at 185℃ for 72 hours[1]. The HCN polymer in the same volume of Milli-Q
water was frozen with liquid nitrogen (-196℃), kept in
the freezer (-18℃), and thawed under room temperature.
This freeze-thaw cycle experiment was conducted 10,
20, 30, and 40 cycles. Each experimental products were
filtered to remove the residual formamide and once
dried.
For the scanning transmission x-ray microscopy(STXM) measurements, each experimental products
were suspended in methanol and dropped in SiOsupported Cu transmission electron microscopy (TEM)
grids. C- and N-XANES spectra of the samples wereacquired by STXM-XANES, beamline 4U, UVSOR.
Cody et al. (2008) [2] was referred to for the absorption
peak assignment.
Figure 1 shows the C-XANES spectra of the products
obtained from the freeze-thaw cycle experiments of
HCN polymer. The spectrum of the starting material (0cycle) contains the developed peak derived from nitrile
or N-heterocycles with the other three peaks derived
from aromatic carbon, carbonyls, and alcohol/ether.
Except for the freeze-thaw 10 cycle product showing
the high peaks of aliphatic and carbonyl carbons, thespectra of the other products are relatively similar tothat of the starting material, but peak intensities ofnitrile /N-heterocycles and carbonyls in the spectra of
the freeze-thaw 30 and 40 cycles products are lower
than those in starting material and the 20 cycle products.
The result shows that HCN polymer is likely hydrolyzed
and altered with the progress of low temperature freezethaw processes. In addition, the freeze-thaw 30 and 40
cycles products showed similar C-XANES spectra to
those of ultracarbonaceous Antarctic micrometeorites

Fig. 1. C-XANES spectra of the products obtained
from the freeze-thaw cycle experiments of HCN
polymer (0, 10, 20, 30, and 40 cycles). Peak A: 1s-π*transition for aromatic carbon (C=C*) at 285.6 eV, peak
B: 1s-π* transition for nitrile (C≡N*) or N-heterocycles(C-N*=C), at 286.6 eV, peak C: 1s-3p/s* transition for
aliphatic carbon (CHx) at 287.5 eV, peak D: 1s-π*transition for carbonyl carbon (OR(C*=O)-C) at 288.4–
288.7 eV, and peak E: 1s-3p/s* transition for alcohol or
ether (CHx-OR) at 289.3 eV.
[1] F. Cataldo, G. Patanè and G. Compagnini, J.
Macromol. Sci. Part A 33 (2009) 1039.
[2] G. D. Cody et al., Meteorit. Planet. Sci. 43 Nr 1/2
353.
[3] H. Yabuta, et al., Geochim. Cosmochim. Acta 214(2017) 172.

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Ⅲ-5 Life, Earth and Planetary Sciences

BL4U

Organics in Asteroid Ryugu Samples: Analytical Approach Utilizing Microand Multi Beam Techniques.
M. Ito1, N. Tomioka1, M. Uesugi2, A. Yamaguchi3,4, N. Imae3,4, N. Shirai5, T. Ohigashi4,6,
M. Kimura3, M-C, Liu7, R.C. Greenwood8, K. Uesugi2, A. Nakato9, K. Yogata9, H. Yuzawa6,
Y. Kodama$, A. Tsuchiyama10, M. Yasutake2, R. Findlay8, I.A. Franchi8, J.A. Malley8,
K.A. McCain7, N. Matsuda7, K.D. McKeegan7, K. Hirahara11, A. Takeuchi2, S. Sekimoto12,
I. Sakurai13, I. Okada13, Y. Karouji14, T. Yada9, M. Abe9, T. Usui9, S. Watanabe13 and Y. Tsuda4,91

JAMSTEC Kochi, 2JASRI/SPring-8, 3NIPR, 4SOKENDAI, 5Tokyo Met. Univ.,
6
UVSOR/IMS, 7UCLA, 8Open Univ., 9JAXA/ISAS, 10Ritsumeikan Univ.,
11
Osaka Univ., 12Kyoto Univ., 13Nagoya Univ., 14JAXA/JSEC, $Toyo Corp.

How the Earth gained its water and organics remains
an outstanding question in planetary science. While asmall fraction may have been inherited from the cloudof gas and dust present when the Solar System formed,
most was likely delivered to the inner Solar System
region by water- and carbonaceous-rich materials i.e.,
asteroids, meteorites, interplanetary dust particles andcomets that may be originated beyond the orbit ofJupiter. To better understand the origin of primitive
water- and carbonaceous-rich asteroids, JAXA sent the
Hayabusa2 spacecraft to asteroid Ryugu and returned
surface and subsurface samples from it in December
2020.
In June 2021, following the initial characterization
phase conducted at JAXA curation facility, a subset of
Ryugu materials were the subject of investigations byseparate research teams of six initial analysis and two
curation teams [1-8]. Based on the investigations ofbulk O isotopes, major elemental abundances and theirmineralogical characteristics [1-3,9], the samples
returned from Ryugu have shown that they closely
match the rare, water rich, Ivuna type (CI) chondritic
meteorites.
Analysis of the organic component in Ryugu particles
has been a major theme during this first intensive phaseof research activity. One of the successes of the work
carried out on the Ryugu materials has been theidentification of a clear relationship between coarse
grained phyllosilicates and organics indicating ofwater-mineral-organics interaction under a reducing
environment on the asteroid [1]. This was confirmed by
a STXM (UVSOR-BL4U), NanoSIMS (JAMSTEC,
Kochi) and TEM (JAMSTEC, Kochi) coordinatedmicroanalysis [1]. Approximately 20,000 organic
species consisting of carbon, hydrogen, nitrogen,oxygen, and/or sulfur, and at least 20 amino-acids havebeen identified by using high-sensitivity mass
spectrometry [1,6,7]. Studies of chiral amino acids in
Ryugu have shown that the abundance of the D- andL-enantiomers are approximately equal (D/L = 1),

indicating nonbiological origins [6]. The diversity oforganic matter found in Ryugu samples shows somesimilarities with that previously reported in primitive
carbonaceous chondrite groups [1,6,7].
Isotope analyses of the Ryugu particles have provided
clues to the original provenance of these Ryugu-derived
materials [1,10-12]. This indicates that materials from
Ryugu display similarities to other postulated outer
Solar System materials, including interplanetary dust
particles, returned cometary materials and spectroscopic
observations of comets. These results suggest that
Ryugu, and by inference other type asteroid, could haveformed in outer Solar System regions, such as are now
occupied by Uranus and Neptune. A finding of CO2bearing water in a Ryugu pyrrhotite is also anotherevidence of outer Solar System origin [4]. Ryugu-likeC-type asteroidal bodies may be present in the inner
Solar System as a result of giant planet migration [13].
The success of the Hayabusa2 mission and subsequent
analytical campaign [1-8] demonstrates the importanceof using state-of-the-art analytical techniques wheninvestigating spacecraft returned samples.
[1] M. Ito et al., Nat. Astron. 6 (2022) 1163.
[2] E. Nakamura et al., Proc. Jpn. Acad., Ser. B 98
(2022) 227.
[3] T. Yokoyama et al., Science 379 (2023) eabn7850.
[4] T. Nakamura, et al., Science 379 (2023) eabn8671.
[5] R. Okazaki et al., Science 379 (2023) eabo0431.
[6] H. Naraoka et al., Science 379 (2023) eabn9033.
[7] H. Yabuta et al., Science 379 (2023) eabn9057.
[8] T. Noguchi et al., Nat. Astron. 7 (2023) 170.
[9] R. C. Greenwood et al., Nat. Astron. 7 (2022) 29.
[10] N. Kawasaki et al., Sci. Adv. 8 (2022) eade2067.
[11] D. Nakashima et al., Nat. Commun. 14 (2023)
532.
[12] T. Hopp et al., Sci. Adv. 8 (2022) eadd8141.
[13] A. Morbidelli, et al., Meteorit. Planet. Sci. 35(2000) 1309.

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BL4U

Chemical Morphology of Biomass-Burning Particles Generated fromDiﬀerent Fuels under Varied Burning Conditions
J. Chen1, N. Faure2, Y. Hao3, Z. Kanji1, E. S. Thomson2, S. Wang3 and X. Kong2*
1
Institute for Atmospheric and Climate Science, ETH Zürich, Zurich, 8092, Switzerland
Department of Chemistry and Molecular Biology, University of Gothenburg, SE-412 96 Gothenburg, Sweden
3
Department of Environmental Sciences, College of Urban and Environmental Sciences,
Northwest University, Xi’an 710127, China

2

Biomass burning aerosols (BBA) from fires
worldwide have the potential to greatly disrupt theatmosphere-Earth climate system, as they directly
absorb solar radiation and also indirectly aﬀect theradiative properties of clouds. However, due to theintricate chemical composition of BBA produced froma variety of fuels under diﬀerent burning conditions,the environmental and climatic repercussions of BBA
remain highly uncertain.
In this study, BBA generated from laboratory burning
experiments were examined by scanning transmission
X-ray microscopy (STXM), in order to understand thefactors influencing the chemical morphologies of BBA.
Various types of biomasses (softwood, hardwood,leaves, peat) were burned under controlled conditions,
including flaming and smoldering conditions. The
generated BBA particles were transferred to the aerosolchamber and collected onto the TEM grid for chemicalmorphology analysis by. The STXM measurements
were performed at the BL4U beamline in UVSOR-III[1].
The images of example particles are shown in Fig.1,
including the BBA particles from softwood underflaming condition (Fig.1a), and softwood, peat, leaves
BBA particles from smoldering condition (Fig. 1b-1d).
The morphologies of particles varied between burning
conditions: particles formed under flaming condition
exhibit a chain-like structure (Fig. 1a), which is typicalfor fresh soot particles. As for particles collected under
the smoldering condition (Fig.1b-1d), they display acore-shell structure, implying a mixture of organics andsoot particles.
The carbon K-edge near edge X-ray absorption finestructure (NEXAFS) spectra of individual particles
from diﬀerent cases were shown in Fig. 2, where thecorresponding particles are displayed on the right. All
spectra show the absorbance around 285 eV (arisingfrom the C 1s→π*R(C*=C)R transition) and a boarder peakat 290 eV (attributing by C 1s→σ* aromatic C=C
transitions). This is one of the identifying features ofsoot particles. Thus, the strong absorbance at 285 eV ofwood-burning particles (softwood flaming andsmoldering) indicates the origin of soot particles. This
peak is relatively lower in peat and leaves smolderingcases, implying less contributions from soot particles.
Two specific peaks at 297.1 eV and 299.8 eV due to thetransitions of K L2 2p1/2→σ* and K L2 2p3/2→σ* wereobserved in the leaves smoldering case. The spectrafeature in the energy range from 285 eV to 290 eVvaried with samples, which is associated with oxygen-containing organic functional groups and needs further
investigation.
These results show that the BBA has distinct chemicalmorphology depending on biomass types and burning
conditions, and the SXTM serves as a useful tool tostudy the morphology and chemical natures of these
BBA particles.

Fig. 1. The morphology of BBA particles from burning
diﬀerent biomasses: (a) Softwood burned under flaming
condition; (b-d) softwood, peat and leaves burned
under smoldering conditions, respectively. The usedphoton energy was 300 eV.

Fig. 2. Left: NEXAFS C K-edge spectra of example
particles. Right: (a) softwood burned under flaming
condition; (b-d) softwood, peat and leaves burned
under smoldering conditions, respectively.
[1] T. Ohigashi et al., J. Phys. Conf. Ser., 463 (2013)
012006.

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Ⅲ-5 Life, Earth and Planetary Sciences

BL5B

Heat Resistance of SiC/Mo/Si Multilayer Mirrors at 30.4nm
S. Teramoto 2, K.Inoue1, A.Yamazaki1, T.Matsumoto2, S. Liu2, K. Yoshioka1,2 and I. Yoshikawa1,2
1Development of Earth and Planetary Science, Graduate School of Science,
The University of Tokyo, Tokyo 113-0033, Japan
2
Development of Complexity Science and Engineering, Graduate School of Frontier Sciences,
The University of Tokyo, Chiba 277-8561, Japan

Optical observations of planetary atmospheres are
conducted over a wide range of wavelengths. In
particular, ultraviolet light is useful for obtaining
various information about planetary upper atmospheres.
For example, by detecting the emission of He II (30.4nm), the distribution of plasma surrounding the Earthcan be observed. The behavior of plasma is related tosolar activity and is the key to understanding thephysics and evolution of the environment within thesolar system. However, the problem is that the signalto-noise ratio is small due to the low light intensity in
UV observations.
Therefore, technology using periodic multilayer forthe normal incidence optics in the EUV wavelength
ranges has been developed for many planetary science
missions, such as Nozomi [1], Kaguya, andEQQULEUS [2]. The normal incidence optics isnecessary to determine the distribution of plasma.
However, the problem is that the normal incidence
optics have low reflectance. We can solve this problem
by using multilayer mirrors.
Multilayer mirrors can dramatically improvereflectance due to the interference of reflected light ateach interface. In particular, the development of highreflectance multilayer mirrors for He II radiation at
30.4 nm is progressing. A Mo/Si mirror is widely used
due to its high stability.
Previously, we demonstrated the performance of a
new multilayer mirror consisting of 40 pairs of Mg and
SiC, achieving a maximum reflectivity of over 30%.
However, it was found that the Mg/SiC mirror had acritical flaw. The reflectivity decreases in hightemperature and high-humidity environments. In actual
observations, multilayer mirrors are exposed to hightemperature and humid environments before launchand extremely cold environments in space. Therefore,
it is necessary to develop observation equipment that is
resistant to such environmental changes.
Since a combination of Mo/Si is stable [3], we
designed a new multilayer mirror (SiC/Mo/Si). The
new mirror is manufactured by depositing the three
diﬀerent materials periodically. To evaluate its
resistance to high-temperature environments, we

conducted a heat environment test in a constanttemperature oven (50 degrees Celsius for 24 hours) andcompared the reflectances of the mirrors before andafter heating.
The synchrotron radiation was spectroscopic with
G3M5. It was led to the calibration chamber. EUV light
was irradiated onto the multilayer mirror, and its
wavelength dependence was evaluated. The
measurement was conducted three times.
Fig. 1 shows the reflectances for the S-polarization of
the mirror before and after high-temperature exposure(50 degrees Celsius for 24 hours). The reflectancedecreased by approximately 5% after the hightemperature exposure. The irregularity of the interface
may have aﬀected the result.
In the future, changing the manufacturing process
will make it possible to create a more stable mirror withhigher reflectance. We plan to improve the performanceof the mirror that is more resistant to environmental
changes and have higher reflectance in the next year.

Fig. 1. Reflectances of SiC/Mo/Si mirrors before(black dots) and after (orange dots) heat-temperature
exposure.
[1] I. Yoshikawa et al., Earth Planets Space 60 (2008)407.
[2] M. Kuwabara et al., JAXA Research andDevelopment Report (2019).
[3] L. Yunpeng et al., Appl. Surf. Sci. 317 (2014) 902.

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BL6B

Synchrotron-radiation Infrared Microspectroscopy Analysis of Persistent
Organic Compounds
T. Kawasaki 1, F. Teshima2 and K. Tanaka2
1

Accelerator Laboratory, High Energy Accelerator Research Organization, Tsukuba 305-0801, Japan
2
UVSOR Synchrotron Facility, Institute for Molecular Science, Okazaki 444-8585, Japan

It is important for absorption of carbon dioxide underatmosphere to manage woody biomass properly. Lignin
is a part of ingredients of woods and is composed ofmany phenolic compounds. It has been known thatthose aromatic molecules such as coumaryl alcohol andconiferyl alcohol are useful for functional materials.
However, it is usually diﬃcult to degrade the lignin
construct unless using strong acids with extremely hightemperatures. We have found that soluble lignin thatwas prepared by alkaline treatment can be degraded to
produce several aromatic molecules by using infrared
free electron laser (IR-FEL) [1]. In this study, an
insoluble sulfonated lignin was tested as another model
sample for degradation by IR-FEL and far-infrared ray
from gyrotron [2]. After those irradiations, the complex
structure of the lignin was analyzed by usingsynchrotron-radiation infrared microspectroscopy. The
measurement was performed by reflection mode with
64 scans.
In the FT-IR spectra of lignin in the far-infrared
region (Fig. 1, upper), increases of absorption intensity
can be observed from 150 to 250 cm-1 in all of samples.
The inclinations in the cases of samples after the IRFEL irradiations at 7.1 μm and 3.5 μm are similar withthe case of the sample before irradiation (Nonirradiation). On the other hand, the irradiation by asubmillimeter wave at 1.4 mm from gyrotron gaveincrease of absorption at 150-200 cm-1. This may
indicate that the hydration of the lignin sample can be
occurred by the irradiation at longer wavelengths.
In the absorption spectra at mid-infrared region (Fig.
1, bottom), broad bands at 1000-1100 cm-1 can beobserved in all of samples. Especially, another peak canbe slightly detected after irradiations at 1.4 mm and 7.1
μm compared to that at 3.5 μm and the non-irradiation
sample (dotted line). This region contains carbonoxygen stretching vibrational modes, and the lignin isconstructed by many ether bonds (C-O-C). Therefore,
it can be considered that the structure of lignin can be
modified and partial cleavage of the ether bonds can be
induced by the submillimeter wave and the IR-FEL
irradiations. We are planning to perform mass analysis
to confirm the structural modification of the lignin.

Fig. 1. Infrared absorption spectra of lignin in the far(upper) and mid-(bottom) infrared regions. “1.4 mm”
means the wavelength of submillimeter wave fromgyrotron. Both of “7.1 μm” and “3.5 μm” arewavelengths of IR-FELs.
[1] T. Kawasaki et al., Polymers 14 (2022) 2401.
[2] T. Kawasaki et al., Biomolecules 12 (2022) 1326.

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Ⅲ-5 Life, Earth and Planetary Sciences

BL6B

Development of an Infrared Spectro-Microtomography Using a Mid-IR
Supercontinuum Laser: Preliminary Report
H. Yabuta1, Y. Ikemoto2, F. Teshima3 and K. Tanaka3
1

Department of Earth and Planetary Systems Science, Hiroshima University, Kagamiyama 1-3-1,
Hiroshima 739-8526 Japan
2
Japan Synchrotron Radiation Research Institute (JASRI) 1-1-1,
Kouto, Sayo-cho, Sayo-gun, Hyogo 679-5198 Japan
3
UVSOR Synchrotron Facility, Institute for Molecular Science, Okazaki 444-8585, Japan

This study aims to develop a three-dimensional
infrared spectro-microtomography using a mid-infrared
supercontinuum laser, for investigating the threedimensional spatial distributions of micron-sizedorganic materials and hydrous minerals of
extraterrestrial samples and for understanding thechemical history in the early Solar System.

In the previous beamtime, the mapping measurements
were performed in the multipoint measurement modeof microscopy programs, but there is a problem of not
being able to access integral intensity plots. In addition,
the program is not equipped with a mode to extract only
the part of the linear array detector that are illuminatedby IR light. The element shift mapping software solves
these problems and is used for the measurement.

In the previous beamtime, an alignment procedure
has been established to guide the laser source to themicroscope for micro-spectroscopy. The activities ofthis beamtime were the following three points. The firstwas to adjust the beam size at the microscope focal
point, the second was to confirm the sample holding
technique, and the third was to confirm the IR imaging
measurement technique by rotating the sample. The
details are described in this order.
Fig. 1 shows the schematic illustration of the optical
path. The IR beam emitted from the laser are reflectedby BaF2 and reduced to about 1/100 of its original
intensity. The IR beam is further attenuated through an
ND filter to an intensity that does not damage theoptical components of the FTIR spectrometer, and thena spectrum is measured. The microscope at BL6B has
16 elements HgCdTe (MCT) linear array detector. In
previous experiments, it was found that due to the small
source size, the IR spot on the microscope was small
and the IR light was only on a part of the element. In
order to increase the number of elements that are
illuminated by IR light, a beam expander was installedin front of the FTIR to adjust the spot size.
Samples are glued to the tip of a glass capillary andmounted on a goniostage. Figure 2(a) shows aphotograph of the goniostage mounted on the
microscope. Figure 2(b) shows a micrograph of asample glued to the tip of the capillary. The sample wasa milled breadcrumb and the size was approximately 90
micrometers in longitudinal.

Fig. 1. Schematic illustration of the optical path

Fig. 2. (a) Photograph of the goniostage mounted on
the microscope. (b) Micrograph of a sample glued tothe tip of the capillary.

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