Ⅲ-5

Life, Earth andPlanetary SciencesⅢ -5. Life, Earth and Planetary Sciences
BL1U
BL1U

Optical Activity Emergence in Amino-Acid Films by Vacuum-Ultraviolet
Circularly-Polarized Light Irradiation (I) - Experimental Setup J. Takahashi1, K. Matsuo2, Y. Izumi2, K. Kobayashi1, M. Fujimoto3 and M. Katoh3
Fucalty of Engineering, Yokohama National University, Yokohama 250-8501, Japan
2
Hiroshima Synchrotron Radiation Center, Higashi-Hiroshima 739-0046, Japan
3
UVSOR Synchrotron Facility, Institute for Molecular Science, Okazaki 444-8585, Japan1

The origin of homochirality in terrestrial bioorganic
compounds (dominant L-amino acid and D-sugar) is
one of the most mysterious issues that remainunresolved in the study of the origin of life. Because
bioorganic compounds synthesized in abiotic
circumstances are intrinsically racemic mixtures of
equal amounts of L- and D-bodies, it is hypothesizedthat chiral products originated from asymmetricchemical reactions. These types of asymmetric
reactions could have possibly been derived fromphysically asymmetric excitation sources in space, thatis “chiral radiation”, and the chiral products would havebeen transported to primitive Earth resulting interrestrial biological homochirality (Cosmic Scenario)[1]. Eventually, terrestrial observations of circularlypolarized light (CPL) radiation due to scattering byinterstellar dust clouds in star formation regions havebeen reported [2].
Several ground experiments have already examinedasymmetric photochemical reactions in simple
biochemical molecules by CPL. We reported opticalactivity emergence in solid-phase films of racemic
mixtures of amino acids by CPL irradiation of 215 nm
in wavelength from free electron laser (FEL) of
UVSOR-II [3].
Recently, we are carrying out the measurement of
CPL irradiation wavelength dependence of opticalactivity emergence by tuning undurator gap conditions.
We have already also reported the optical activity
emergence by using CPL irradiation of 230, 215, and203 nm in wavelength from undulator BL1U of
UVSOR-III [4].
Presently, we are trying to introduce optical activity
by vacuum ultraviolet (VUV) CPL irradiation in shorter
wavelength than 200 nm . As for the sample, we formed
thin solid films of racemic DL-alanine on fused quartz
substrates using a thermal-crucible vacuum evaporator
from crystal powders of DL-alanine as a sublimation
source. Sublimation temperature was in the range of150~200°C and pressure of the vacuum chamber wasapproximately 10-2 Pa throughout the evaporation
process. In order to introduce optical activity into theracemic film, we irradiate them with CPL introduced
from undulator BL1U of UVSOR-III. The samples
were set in a vacuum sample chamber preventing VUVCPL attenuation by air absorption in shorter wavelengththan 200 nm (Fig.1). On the beam entrance side of thevacuum sample chamber, a gate valve with a vacuumsealed MgF2 window was mounted. The irradiated CPL

wavelengths were 180, 155, and 120 nm corresponding
to photon absorption bands of alanine molecule. The
irradiated photon number was measured withphotoelectron current of a silicon photodiode(International Radiation Detectors, Inc.) settled at the
sample position. The typical energy dose calibrated byquantum efficiency of the silicon photodiode wasapproximately 50mWhour.
In order to clarify the optical anisotropy of the films
before and after VUV CPL irradiation, we measured
the synchrotron radiation (SR) circular dichroism(CD) at beamline BL-12 of Hiroshima Synchrotron
Radiation Center (HiSOR). In order to delete theeffects of linear dichroism (LD) and/or linear
birefringence (LB) components, dependence on
sample rotation angle (0, 45, 90, and 135degrees) of
the CD spectra was measured.
Through the VUV CPL irradiation experiments andthe detailed analysis of SR-CD spectra, we are aiming
to clarify full mechanism of the optical activity
emergence, which potentially has relevance to theorigin of terrestrial biological homochirality.

Ⅲ- 5Fig. 1. Photo of VUV CPL irradiation experiment on
BL1U using a vacuum sample chamber.
[1] W. A. Bonner, Orig. Life Evol. Biosph. 21 (1991)
59.
[2] T. Fukue et al., Orig. Life Evol. Biosph. 40 (2010)335.
[3] J. Takahashi et al., Int. J. Mol. Sci. 10 (2009) 3044.
[4] K. Matsuo et al., UVSOR Activity Report 44(2017) 157.

1471 Ⅲ05-

Ⅲ -5. Life, Earth and Planetary Sciences
BL1B, BL7B
BL1B, BL7B

Giant Thermal Effect of Vibration Modes of Single-Crystalline Alanine
2Ⅲ- 5

Z. Mita1, H. Watanabe1, 2 and S. Kimura1, 2

Graduate School of Frontier Biosciences, Osaka University, Suita, 565-0871, Japan
Department of Physics, Graduate School of Science, Osaka University, Toyonaka, 560-0043, Japan
1

Amino acids are the most basic molecules of living
bodies. Since almost all amino acids are chiralmaterials, they have enantiomers, L- and D-form.
Although amino acids are well-known, fundamental
physical properties have hardly been studied. Owing
to the crystal structure, strong anisotropy of opticalconstants and ferroelectricity are expected. However,
the optical measurements using single-crystalline
samples have never been studied so far, neither thetemperature dependence. Here we report thetemperature-dependent anisotropic reflectivity spectraof single-crystalline L-alanine that has the simplest
molecular structure among amino acids havingchirality.
We have grown the single crystal by using thesolvent evaporation method and confirmed to beorthorhombic structure reported previously [1].
Polarized optical spectra from the terahertz (THz) tovacuum-ultraviolet (VUV) regions have beenmeasured by using BL1B [2] and BL7B [3] at
UVSOR-III and laboratorial equipments. Temperaturedependence of anisotropic reflectivity spectra in theTHz region are shown in Fig. 1. Some peaks owing tomolecular vibrations appear. The optical spectra arestrongly anisotropic and all of these peaks have large
temperature dependency. For instance, the intensity ofthe peak appearing at the photon energy of about 15
meV (indicated by a red square in Fig. 1) in theconfiguration of the electric vector parallel to the caxis (E // c) increases 4 times larger with decreasing
temperature from 300 K to 10 K. The obtainedreflectivity spectra were converted to the opticalconductivity spectra by using the Kramers-Kronig
analysis. The temperature-dependent peak at 15 meV
is shown in Fig. 2.
To explain the temperature dependence, we adopt
the Morse potential that is one of the simplified
potentials having anharmonicity [4]. Here, we assume
the Boltzmann distribution for oscillators’ occupation.
Assuming the multiple excitation among energy
levels, the temperature dependence of the peak in Fig.
2 can be reproduced by using the summation ofLorentz functions owing to the excitations. The results
reproduce well as shown by thin lines in Fig. 2.
Furthermore, using the parameters obtained from thesimulation, the bottom energy of the Morse potential
can be evaluated as 822 meV at 10 K and 675 meV at
300 K. The change of the potential shape is the originof the giant thermal effect of the molecular vibrationsof single-crystalline L-alanine.

Fig. 1. Anisotropic reflectivity spectra at temperaturesfrom 10 to 300 K in THz regions perpendicular (a) andparallel (b) to the c-axis.

Fig. 2. Temperature-dependent optical conductivity[()] spectra of the 15meV peak (thick lines). Thinlines are the simulation results using a simplified
anharmonic Morse potential.
[1] M. Fleck and A. M. Petrosyan, Salts of Amino
Acids-Crystallization Structure and Properties(Springer, 2014) p. 26.
[2] S. Kimura, E. Nakamura, K. Imura, M. Hosaka, T.
Takahashi, M. Katoh, J. Phys.: Conf. Ser. 359 (2012)
012009.
[3] K. Fukui, H.Miura, H. Nakagawa, I. Shimoyama,
K. Nakagawa, H. Okamura, T. Nanba, M. Hasumoto,T. Kinoshita, Nucl. Instrum. Meth. Phys. Res. A 467468 (2001) 601.
[4] P. M. Morse, Phys. Rev. 34 (1929) 57.

1482 Ⅲ05-

Ⅲ -5. Life, Earth and Planetary Sciences
BL3U
BL3U

Novel Insights to Cloud Water Microphysics Using Synchrotron-excitedXAS
J. J. Lin1, G. Michailoudi1, H. Yuzawa2, H. Iwayama2, 3, M. Nagasaka2, 3,
M. Huttula1, N. Kosugi2, 3 and N. L. Prisle1

Nano and Molecular Systems Research Unit, University of Oulu, Oulu 90041, Finland
UVSOR Synchrotron Facility, Institute for Molecular Science, Okazaki 444-8585, Japan3
School of Physical Sciences, The Graduate University for Advanced Studies (SOKENDAI), Okazaki 444-8585,
Japan
1

2

Cloud droplets in the atmosphere are complex
aqueous mixtures of organic and inorganic
constituents, but their exact composition and thenature of aqueous phase molecular interactions are
extremely challenging to quantify [1]. Amphiphiliccompounds present in the organic aerosol fraction are
known to self assemble into micelles, but direct
chemical information on the aqueous phaseinteractions of amphiphilic monomers and theiraggregate structures has been lacking.
Two types of samples were analyzed withsynchrotron radiation X-ray absorption spectroscopyusing the liquid cell setup at BL3U [2]: 1) cloud watercollected from October 10-14 at the Finnish
Meteorological Institute Pallas-Sodankylä Global
Atmosphere Watch station in the sub-Arctic region in
Finnish Lapland as part of the 7th Pallas CloudExperiment from September 1 to November 30; and 2)
organic micellar systems composed of n-octanoic acid,n-decanoic acid, and sodium n-decanoate dissolved inwater at varying multiples of their critical micelleconcentration (CMC). A summary of the samples
measured is given in Table 1.
For the cloud water samples, both the oxygen Kedge in the 528-550 eV range and the carbon K-edge
in the 280-300 eV range were studied. Unfortunately,
due to the apparently very low concentration oforganic material in the cloud water, presumably fromvery clean Arctic conditions, any difference between
cloud water spectra and pure water spectra could not
be detected. Another batch of the cloud water sample
will be subject to various complementary analyses toconfirm this.
For the aqueous organic micellar solutions, thecarbon K-edge in the 280-300 eV range was studied.
Photon energies in the 280-285 and 290-300 eV range
were scanned at 0.1 eV resolution while photon
energies in the 285-290 eV range were scanned at 0.02
eV resolution in order to specifically capture thealiphatic R(CH)R’ and carboxylic C(=O)OH carbon
absorption peaks from monomers to identify shifts inpeak location due to the difference in chemical
environment between monomers and micelles insolution.
Preliminary absorption spectra at the C 1s
absorption edge for sodium n-octanoate in water atvarious concentrations along with pure octanoic acidare shown in Fig. 1. Background absorption due towater and the cell membranes have been removed. For
all the samples, one main peak is observed at around289 eV. For sodium n-octanoate solution below the
CMC, the absorption spectra before and after the mainfeature are less than those solutions above the CMC.
Interestingly, the higher the sodium n-octanoate
concentration in solution, the more the absorption
spectra start to resemble that of pure octanoic acid. The
measurements hold promise for detecting the aqueousphase interactions of organic monomer and micellar
structures compounds in solution.
Table 1. Summary of samples measured
SampleCloud Water
n-octanoic
acid
Sodiumn-octanoate
Sodiumn-decanoate

CMC
[mM]
–
140. [3]340.0 [4]94.0 [4]Concentrations–
0.75, 1.25, 2, 3, 4, 6,
and 8 times CMC;
pure
0.75, 2, 3, 4, and 5
times CMC
2, 3, 4, 6, and 8 times
CMC

Numberof scans
5
15
7
10

Ⅲ- 5

Fig. 1. Preliminary absorption spectra for sodium noctanoate solutions at various concentrations together
with that of pure octanoic acid.
[1] T. Kurtén et al., J. Phys. Chem. A 119 (2014) 4509.
[2] M. Nagasaka et al., J. Electron Spectrosc. 177(2010) 130.
[3]A. N. Campbell and G. R. Lakshminarayanan, Can.
J. Chem. 43 (1965) 1729.
[4] K. Quast, Miner. Eng. 19 (2010) 582.

1493 Ⅲ05-

Ⅲ -5. Life, Earth and Planetary Sciences
BL4U
BL4U

Feasibility Study of Sulfur Speciation High-spatial Resolution Mapping in
Extraterrestrial Organics by STXM-XANES
M. Ito1, R. Nakada1, H. Suga2, T. Ohigashi3, Y. Kodama4 and H. Naraoka5

Kochi inst. for Core Sample Res., JAMSTEC, Nankoku 783-8502, Japan
2
Hiroshima University, Higashihiroshima 739-0046, Japan
3
UVSOR Synchrotron Facility, Institute for Molecular Science, Okazaki 444-8585, Japan
4
Marine Works Japan LTD., Japan
5
Kyushu University, Fukuoka 819-0395, Japan1

Ⅲ- 5

Sulfur is one of the major elements in terrestrial andextraterrestrial organics. The elemental compositions of
the Murchison IOM (insoluble organic matter) areproposed to be C100H70O22N3S7 [1] or C100H48N1.8O12S2 [2].
Because sulfur shows a wide range in oxidation state (-2 to+6) with both electropositive and electronegative elements,
reduced and oxidized sulfur species have been found invarious carbonaceous chondrites [3, 4]. Therefore,
understanding of speciation of sulfur and its distribution
within organics in a carbonaceous chondrite may provide
the secondary alteration processes of thermal
metamorphism and aqueous alteration in the parent body.
Synchrotron based X-ray absorption near edge
spectroscopy (XANES) is a powerful analytical tool tocharacterize and quantify chemical speciation, functionalgroup and bonding environment of the sample. In the fieldof cosmochemistry, many researches were carried out toidentify functional groups of C, N and O in extraterrestrialorganics (i.e., IOM in carbonaceous chondrites [5],cometary returned sample [6], organics found in Hayabusa
Category-3 particles [7], organics in IDPs [8], and organic
component extracted from halite grain in Monahans LL
chondrite [9]). However, sulfur study using XANES in theextraterrestrial organics is very rare [3, 10].
In this study, we report preliminary results of sulfur
speciation measurements by L3-edge XANES of series
sulfur bearing terrestrial organics to obtain reference
spectra, and of S, N, C and O-XANES in the FIB sectionof the Murchion CM2 carbonaceous chondrite (147 x 187
pixels, 22 x 28 µm2: spatial resolution = 150 nm) using
scanning transmission X-ray microscope (STXM) at Inst.
Mole Sci. UVSOR BL4U. We also took a sulfur image(L3-edge XANES) of organics extracted from
Asuka881458 CM2 chondrite for comparison in terms ofsulfur distribution and speciation.
We show nine reference sulfur L3-edge spectra
measured by the STXM. Reference sulfur bearingorganics are sodium lauryl sulfate, sodium
methanesulfonate, dibenzothiophene, thianthrene, DLmethionie, DL-methionine sulfone, L-cysteic acid, Lcystein, and L-cystine (Fig. 1). These organics show
different absorption curves which related to sulfur related
chemical bond (e.g., sulfate, sulfone, thiol).
We successfully obtained high resolution sulfur
STEXM images of both FIB sections of Murchison andorganics from Asuka881458. We will continue to measureanother primitive chondrite for understanding of diversityof organics based on sulfur speciation. We plan to combine
sulfur isotope imaging with NanoSIMS for same sections
that used in this study.
[1] L. Remusat, EPJ Web of Conferences 18 (2011)
05002.
[2] Gilmour, Meteorites, Comets, and Planets, Treatiseon Geochemistry (2005) 269.
[3] Orthous-Daunay et al., EPSL 300 (2010) 321.
[4] W. J. Cooper et al., Science 277 (1997) 1072.
[5] G. D. Cody et al., Meteor Planet Sci. 43 (2011) 353.
[6] S. A. Sanford et al., Science 314 (2006) 1720.
[7] H. Yabuta et al., EPS 66 (2014) 156.
[8] G. J. Flynn et al., Geochim Cosmochim Acta 67(2003) 4791.
[9] Chan et al., Science Advance 4 (2018) eaao3521.
[10] M. Bose et al., Meteor Planet Sci. 52 (2017) 546.

Fig. 1. Sulfur bearing organics of L3-edge STXM-XANES spectrum

1504 Ⅲ05-

Ⅲ -5. Life, Earth and Planetary Sciences
BL4U
BL4U

Organic Vein in a Primitive Clast in the Carancas Meteorites
Y. Kebukawa1, M. E. Zolensky2, T. Ohigashi3 and Y. Inagaki3

Faculty of Engineering, Yokohama National University, Yokohama 240-8501, Japan
2
ARES, NASA Johnson Space Center, Houston, TX 77058, USA
3
UVSOR Synchrotron Facility, Institute for Molecular Science, Okazaki 444-8585, Japan1

Primitive xenolithic clasts are often found in various
groups of meteorites [e.g., 1, 2]. These xenoliths
sometime contain unique carbonaceous matter. We
have been analyzed these using scanning transmission
X-ray microscopes (STXM) combined with othermicroscopic techniques [3-5]. Some of these could becandidates of primordial trans- Neptunian object(TNOs) that have been scattered by giant planetary
migration. It was suggested that numerous bodies was
injected from outer Solar System into the main
asteroid belt, where modeling shows they can
successfully reproduce the observed D/P-type asteroid
populations [e.g., 6]. After implantation, some of the
D/P-asteroids would have collided with parent body(s)of ordinary chondrites (S-type asteroids). Some of thefragments from D/P-asteroids left in the ordinarychondrite parent body(s) and eventually could bedelivered to Earth as meteorites [7]. The Carancas
meteorite is a H4-5 ordinary chondrite that contains aprimitive xenolithic clast rich in organic matter.
We are conducting coordinated analysis of organic
matter in the clast using various methods [8]. 100 nmthick FIB sections were prepared from the clast usinga Quanta 3d field-emission gun FIB instrument at
NASA JSC. We investigated the FIB sections with
C,N,O-X-ray
absorption
near-edge
structure(XANES) using the STXM at BL4U of UVSOR.
Figure 1 shows a carbon map containing ~10 µm Crich vein in the clast from Carancas meteorite. CXANES spectrum of the C-rich vein shows 285.0 eV
absorption designed to aromatic carbon and 286.5 eVabsorption designed to ketone (C=O), as well as a
weak absorption at 288.7 eV assigned tocarboxyl/ester [(C=O)O] (Fig. 2). The N-XANES
shows absorption at 398.6, 399.7 and 401.1 eV. The
398.6 and 399.7 eV can be attributed toimine/nitrile/pyridine groups, and 401.1 eV can be
attributed to amide/pyrrole groups [e.g., 9].
The C-XANES features confirmed that the clast was
not subjected to thermal metamorphism in the
Carancas H4-5 parent body. The C-XANES of theorganic vein is distinguished from that of typicalprimitive chondritic insoluble organic matter (IOM),
and dominated in C=O structures. It indicates that theorganic matter is originated in relatively oxygen richcondition. The vein-like morphology indicates thatfluid was involved during formation of this structure.
Thus the organic vein is formed during aqueousalteration in the clast parent body.

Fig. 1. Carbon map obtained from a FIB section fromthe clast in the Carancas meteorite. The while area isorganic vein-like structure. The map was derived with
292 eV and 280 eV images. The scale bar is 5 μm.

Ⅲ- 5Fig. 2. The C-XANES spectrum obtained from theorganic vein in the clast from the Carancas meteorite.
[1] A. J. Brearley, Geochim. Cosmochim. Acta 54(1990) 831.
[2] M. Zolensky et al., Meteoritics 27 (1992) 596.
[3] Y. Kebukawa et al., Geochim. Cosmochim. Acta
196 (2017) 74.
[4] Q. H. S. Chan et al., Sci. Adv. 4 (2018) eaao3521.
[5] Y. Kebukawa et al., Nat. Astro. Submitted.
[6] H. F. Levison et al., Nature 460 (2009) 364.
[7] M. Zolensky et al., LPSC abst. (2018) 1789.
[8] Q. H. S. Chan et al., LPSC abst. (2018) 1191.
[9]P. Leinweber et al., J. Synchrotr. rad. 14 (2007) 500.

1515 Ⅲ05-

Ⅲ -5. Life, Earth and Planetary Sciences
BL4U
BL4U

Direct Observation of the Basalt-cell Interface by STXM-Study on the Mechanism of Microbial Alteration of Oceanic CrustS. Mitsunobu1, 2, Y. Ohashi2 and T. Ohigashi3

Department of Agriculture, Ehime University, Matsuyama 790-8566, Japan
Institute for Environmental Sciences, University of Shizuoka, Shizuoka 422-8526, Japan
3
UVSOR Synchrotron Facility, Institute for Molecular Science, Okazaki 444-8585, Japan
1

2

Ⅲ- 5

Alteration of oceanic basalts is an important waterrock interaction ubiquitously occurring in the vast
oceanic crust. The alteration of oceanic basalt has asignificant impact on Earth’s climate on a geologicaltimescale by providing a sink for atmospheric CO2
through carbonization of oceanic basalts (e.g.,Standigel et al., 1989; Sleep & Zahnle, 2001). The
fresh basalt rock contains abundant Fe (roughly 9wt.%) and its dominant species is ferrous iron (Fe(II))
and Fe(II)/Fetotal ratio in fresh basalt ranges between
82 and 90% [1]. Hence, when the oceanic basalts are
exposed to oxidative conditions (e.g., oxygensaturated seafloor), the Fe(II) in basalts is gradually
oxidized to Fe(III) in the alteration process on ageological timescale even at low temperature in deep
seafloor [2]. Recent studies have noticed that oceanic
microbes (mainly bacteria and archaea) play asignificant role to the oxidation of Fe(II) in the basalt.
Also, microbial Fe(II) oxidation accelerate the wholealteration process of oceanic crust [2], as the rate ofbiogenic Fe(II) oxidation is generally more rapid thanthat of abiotic (chemical) oxidation. There have been
considerable efforts to identify mechanisms of theoxidation of Fe(II) involved in the oceanic basalt.
However, little is known on the mechanism of
microbial Fe(II) in the basalt alteration, because direct
chemical speciation of metals and biomolecules atmineral-microbe interface has been difficult due tohigh spatial resolution in analysis.
Here, we investigated the mechanisms of the Fe(II)oxidation in basalt alteration by scanning transmission
X-ray microscopy (STXM) based C and Fe near edge
X-ray absorption fine structure (NEXAFS) analyses at
UVSOR BL4U. The basalt samples used in the study
was the basalt glass samples incubated for 12 months
at deep seafloor (the depth of waster: ca. 700 m) in IzuOgasawara bonin.
Figure 1 shows the STXM-based merged Fe/C
image and C 1s NEXAFS. The C NEXAFS showedthat the spectrum measured at bacteria cell-basalt
interface in the incubated basalt glass contains asignificant peak specific for acidic polysaccharides
like alginate, addition to the other biomolecules(protein , DNA, and lipid), and this spectral feature
was not found in those of the other area. This finding
indicates that the bacteria on the basalt glass produce
the polysaccharide-rich extracellular polymeric
substances (EPS) at the cell-basalt interface. The
biogenic polysaccharides often form a strong complex

with Fe ion under wide pH region [3]. Thus, ourfinding obtained by C NEXAFS analysis imply thatobserved bacteria could produce the polysaccharides
to enhance the dissolution of basalt by thecomplexation and/or mediate contact between the celland basalt surface. These findings could be important
information for understanding key mechanism ofbiogenic alteration of ocean crust, basalt.

Fig. 1. STXM-based C and Fe images (upper) and C
1s NEXAFS spectra (bottom).
[1] A. Bezos and E. Humler, Geochim. Cosmochim.
Acta 69 (2005) 711.
[2] W. Bach and K.J. Edwards, Geochim. Cosmochim.
Acta 67 (2003) 3871.
[3] S. Mitsunobu et al., Chem. Lett. 44 (2015) 91.

1526 Ⅲ05-

Ⅲ -5. Life, Earth and Planetary Sciences
BL4U
BL4U

3-Dimensional Observation of a Cell Nucleus by Using a Scanning
Transmission X-ray Microscope
T. Ohigashi1, 2, A. Ito3, K. Shinohara3, S. Tone4, Y. Inagaki1 and N. Kosugi1, 2

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
3
School of Engineering, Tokai University, Hiratsuka, 259−1292, Japan
4
Graduate School of Advanced Science and Technology, Tokyo Denki University, Hatoyama, 350-0394, Japan1

2

back-projection algorithm. The reconstructed 3D
distribution of DNA and 2D distributions of DNA andprotein are shown in Fig. 2. While the 2D distributionof DNA in Fig. 2(b) shows distinct sub-nuclear
structures (shown by arrows), presumably nucleoli,that of protein in Fig. 2(c) shows only the compartment
like structures and less protein in the nucleoli.
OpticalDensity[a.u.]

Apoptosis is a process of a programmed cell death
observed in many biological processes. Nuclear
condensation and DNA fragmentation occur duringexecution of apoptosis and this process is classified
into 3 stages. Generally, apoptosis is observed by using
fluorescence microscope with staining process but
high resolution image, chemical compositions and 3dimensional (3D) structure are still unclear. Then, we
use a scanning transmission X-ray microscopy(STXM). In these years, we have been developing
computed tomography (CT) to perform 3D
spectroscopy by changing the energy of the X-ray [1,2]. By using this technique, 3D distributions of protein
and DNA of an isolated cell nucleus were observed.
As the sample preparation, isolated nuclei fromHeLa S3 cells were attached on a slide glass using celladhesive agent, Cell-Tak (Fisher Scientific), and were
fixed by glutaraldehyde. After critical point drying
process, the isolated cell nucleus was glued on a tip ofa tungsten needle. The sample was mounted on a CT
sample cell.
Experiment was performed at BL4U. To perform
CT, a focal depth of a Fresnel zone plate (FZP) should
be longer than thickness of the sample to assume pathsof the X-rays through the sample as a parallel beam.
Therefore, the FZP with smaller NA was used insteadof higher spatial resolution. In this experiment, theFZP with the outermost zone width of 45 nm was used
and the focal depth of 5.2 µm at 400 eV is nearlyenough length for a cell nucleus.
NEXAFS spectra of DNA and histone as protein
around N K-edge are shown in Fig.1. These spectra
show remarkable difference on 1s-* transition so thatthey were used as reference spectra. In CT
measurement, 41 X-ray transmission images (energy
stack) were acquired by changing the X-ray energy
from 397 to 410 eV. 50 energy stacks were acquired byrotating the sample 3.6 each (180 rotation in total).
Then, a size of one image was 88 µm2 with 5050
pixels and a dwell time was 2 ms per a pixel.
For reconstruction of 3D chemical distributions, the
X-ray transmission images of the energy stack werealigned and 2D distributions of DNA and protein wereobtained by fitting their reference spectra to eachenergy stack by using aXis2000 software. Sinograms
were extracted from data sets of the 2D distributionsof DNA and protein respectively and cross sectional
images were reconstructed by home-made filtered

395

400

405

410

Energy [eV]

415

420

Fig. 1. NEXAFS spectra of DNA (blue) and protein(red) at N K-edge.

(a)
Ⅲ- 5

(b)

(c)

2 µm

Fig. 2. (a) 3D volume projection image ofdistribution of DNA and 2D distribution of (b) DNA
and (c) protein at the dashed line in (a).
[1] T. Ohigashi, Y. Inagaki, A. Ito, K. Shinohara and N.
Kosugi, UVSOR Activity Report 44 (2017) 39.
[2] T. Ohigashi, Y. Inagaki, A. Ito, K. Shinohara and N.
Kosugi, J. Phys.: Conf. Ser. 849 (2017) 012044.

1537 Ⅲ05-

Ⅲ -5. Life, Earth and Planetary Sciences
BL4U
BL4U

Investigation of Carbonaceous Materials Inside Moderately Shocked
Ureilite Using STXM-XANES System2

Ⅲ- 5

M. Uesugi1, T. Ohigashi2 and Y. Inagaki2

Japan Synchrotron Radiation Research Institute, JASRI, Sayo 679-5198, JapanUVSOR Synchrotron Facility, Institute for Molecular Science, Okazaki 444-8585, Japan, Japan
1

Ureilite is a unique class of meteorite which shows
small carbonaceous materials inside veins orsometime inclusion like structure, inside the rocky
matrix [1-3]. It shows iron-sulfide inclusions or veins,
indicating that the meteorite would not come fromdifferentiated asteroids (Fig. 1). On the other hand,silicate materials inside the ureilite shows igneous
texture, and their composition is similar to peridotite
of Earth, those formed from highly differentiatedmantle. On the other hand, oxygen isotopic
composition of the ureilite shows close relation withthe carbonaceous chondrites, most primitive class of
meteorites. In addition, existence of carbonaceous
materials also indicates their relation. However,abundance of the carbonaceous materials inside theureilite is much higher than carbonaceous chondrites,
and the carbon material includes euhedral graphite and100µm-sized diamond inclusions, those rarely foundin carbonaceous chondrites. It is interesting thatureilites those showing low impact features do notshow diamond inclusions. It might indicate that
diamond inclusions inside the carbonaceous material
would be formed by heavy shock on the asteroids.
However, those carbonaceous material shows noble
gas content. It is unlikely that long-heated and highlyshocked materials contain such abundant noble gas.
Thus, origin of carbonaceous materials inside theureilite is still unknown, though it might haveimportant information for the time series of evolutionof carbonaceous chondrites.
We investigated Kenna meteorite, a ureiite whichshows moderately shocked texture, using x-ray
diffraction computed tomography (XRD-CT) at
SPring-8, in order to investigate the relation between
the shock degree and characteristics of carbonaceous
materials. We found that distribution of several
different phases of carbonaceous materials inside themeteorite, those are not reported previously.
In this study, we investigated the carbonaceous
materials inside Kenna meteorite using scanningtransmitted x-ray microscope and x-ray absorption
near edge analysis (STXM-XANES), to investigate
and identify the phases of carbonaceous materials
included inside the meteorite. We lifted up several
ultrathin-sections from several points of the meteorite
using Focused ion beam (FIB), and investigated thedifference of the molecular structure of the carbon
phases. Figure 2 shows the result for the STXMXANES analysis of a thin section of carbonaceous
materials picked up from the vein of the meteorite. Itshows quite complicated structure of graphite,diamond, amorphous and some unknown phases.
Unfortunately, we could not find the specific phases
appeared in the XRD-CT image in this investigation
yet. After the analysis of UVSOR, we plan toinvestigate same sections by transmission electron
microscope (TEM) and electron diffraction system, forhigh resolution analysis.

Fig. 1. An X-ray CT image of Kenna meteorite. The
with of the sample is 3mm. White veins are ironsulfide and dark portions are carbonaceous inclusions.
Some of veins also includes carbonaceous material.

Fig. 2. Mapping of carbonaceous materials. (green:
amorphous, red: graphite, blue: diamond, black:
unknown). The width of the section is 20 µm.
[1] J. L. Berkley, G. J. Taylor, K. Kell, G. E. Harlow
and M. Pnnz, Geochim. Cosmochim. Acta, 44 (1980)
1579.
[2] H. Takeda, Earth Planet. Sci. Lett. 81 (1987) 358.
[3] H. Takeda, Earth Planet. Sci. Lett. 93 (1989) 181.

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BL4U

Silica-bearing Minerals in Martian Meteorites
H. Suga1, N. Shiraishi1, N. Sago1, M. Miyahara1, T. Ohigashi2, Y. Inagaki2, A. Yamaguchi3,
N. Tomioka4, Y. Kodama5 and E. Ohtani6

Department of Earth and Planetary Systems Science, Graduate School of Science, Hiroshima University,
Higashi-Hiroshima 739-8526, Japan
2
UVSOR Synchrotron Facility, Institute for Molecular Science, Okazaki 444-8585, Japan
3
National Institute of Polar Research, Tokyo 190-8518, Japan
4
Kochi Institute for Core Sample Research, Japan Agency for Marine-Earth Science and Technology, Nankoku
783-8502, Japan
5
Marine Works Japan, Nankoku 783-8502, Japan
6
Department of Earth Sciences, Graduate School of Science, Tohoku University, Sendai 980-8578, Japan1

Nakhlite is a unique Martian meteorite clan because it
has stark evidence for a rock-fluid reaction occurred on
the Mars. Olivine [(Mg,Fe)2SiO4] occurs in Nakhlite.
One of representative evidence for the rock-fluid reactionin Nakhlite is “iddingsite”, which is the alteration texture
formed in and around the olivine grain. Varied kinds of
secondary minerals occur in the iddingsite [e.g., 1]. The
mineral species, assemblages, compositions andchemical species of the secondary minerals depend on
several parameters such as the temperature and pH of thefluid. Accordingly, Nakhlite allows us to elucidate thephysicochemical properties of the fluid existed on ancientMartian surface and its origin. Many previous studies
propose that sulfate minerals exist on the Martian surface[e.g., 2]. When sulfate minerals dissolve into solution,
sulfate anion and hydrogen ion occur in the solution,
which decreases the pH value in the solution. Silica
precipitates through the dissolution of olivine under very
low pH condition (pH = 0~3); (Mg,Fe)2SiO4 + 4H+ →Si(OH)4 + 2(Mg,Fe)2+. Si LIII-edge XANES allows us toclarify chemical bonding state around Si [2]. However,
Si-XANES was not taken from a natural sample, because
the transmittance of the Si LIII-edge energy (around 100eV) is very low. Hence, we attempt to take Si-XANES
from the iddingsite in Nakhlite.
Nakhlites, the Yamato (Y) 000593 and Yamato (Y)000749 were used for this study. Several portions
including the iddingsite were extracted by FIB. We
prepared the FIB foils having two different thicknesses(Fig. 1). We obtained O-, S- and Fe-XANES from thethick portion, whereas Si- and S-XANES from the thinportion. The foils were observed with TEM/STEM- EDS
after STXM analysis.
Laihunite, ferrihydrite, amorphous or poor crystallized
silica-rich portion and iron sulfate were identified as asecondary mineral in the iddingsite throughTEM/STEM-EDS and STXM analysis. Based on SXANES, sulfur included in the iron sulfate is hexavalent(Fig. 2(b)). Considering TEM/STEM-EDS analysis andselected area electron diffraction (SAED) patterns, theiron sulfate is natrojarosite [NaFe3(SO4)2(OH)6]. Si LIIIedge XANES spectrum were successfully obtained fromthe poor crystallized silica-rich portion besides
montmorillonite and opal as a reference material (Fig.
2(a)). The Si-XANES obtained from the poor crystallized

silica-rich portion appears to be montmorillonite (claymineral: smectite group), which is supported by the
SAED patterns. It is likely that the dissolved Si under low
pH condition precipitates as clay mineral.

Fig. 1. The FIB foil extracted from the iddingsite in Y
000749. a) HAADF-STEM image. The yellow dottedline shows the distribution of the silica-rich portiojn.
b) A shcematic of crossection of the FIB foil.

Fig. 2. Representative XANES spectrum. a) SiXANES obtained from the poor crystallized silicarich portion (blue). Reference spectrum are also shown.
b) S-XANES obtained from the iron sulfate coexistingwith ferrihydrite in Y 000593 (red). Reference
spectrum are also shown.
[1] H. Suga et al., Goldschmidt Conference 2017(2017) 3814pdf.
[2] J. M. Lewis et al., Astrobiology 15 (2015) 247.
[3] L. A. J. Garvie and P. R. Busek, Am. Mineral. 84(1999) 946.

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BL4U

Quantitative Mapping of DNA and Protein in Human Chromosome bySpectromicroscopy with STXM Using Combined NEXAFS Measured at theC, N, and O-K Edges
A. Ito1, K. Shinohara1, T. Ohigashi2, 3, S. Tone4, M. Kado5, Y. Inagaki2 and N. Kosugi2, 3

School of Engineering, Tokai University, Hiratsuka 259-1292, JapanUVSOR Synchrotron Facility, Institute for Molecular Science, Okazaki 444-8585, Japan
3
SOKENDAI (The Graduate University for Advanced Studies), Okazaki 444-8585, Japan
4
Graduate School of Science & Engineering, Tokyo Denki University, Hatoyama 350-0394, Japan
5
Kansai Photon Science Institute, Nat. Inst. Quantum and Radiological Sci. Technol., Kizugawa 619-0215, Japan
1

2

Ⅲ- 5

Spectromicroscopy using scanning transmission Xray microscope (STXM) has been applied to DNA andprotein distributions in biological specimens such as
chromosome and sperm at the C-K absorption edge [1,2]. We proposed the use of a unique NEXAFS peak of
DNA at the N-K absorption edge for DNA imaging [3],and in our previous study using STXM at BL4U, we
obtained DNA and histone, a nuclear protein,
distributions in mammalian cells using NEXAFS at
the N-K absorption edge [4].
In the present study, to extend the usefulness of theunique DNA peak we developed the procedure forquantitative mapping of DNA, protein and otherbiomolecules. In addition, the resultant images werecompared with the results obtained by the widely used
analytical method named as SVD (Singular Value
Decomposition).
We used chromosomes for the mapping of DNA,histone and other biomolecules, because they haverather simple molecular composition, DNA andhistone. Cultured Chinese Hamster Ovary (CHO) cells
were treated with colcemid to stop cell cycle at the
beginning of mitotic phase, then mitotic cells that weredetached from culture dish were collected and fixedwith the mixed solution of ethanol and acetic acid (3:1).
The fixed chromosomes were dropped on formvar
membrane, and then dried in air. For
spectromicroscopy, we measured NEXAFS of DNA,histone, a lipid, phosphatidylcholine (PC), at the C, N,
and O-K edges. The image and spectrum analyses
were performed using aXis2000 software.
Our procedure to obtain molecular maps is asfollows: Images obtained at three absorption edges are
combined to form one image stack file. NEXAFS
spectra are also combined to form one spectrum. DNA
image is obtained by the difference between images on
and below the unique DNA peak at the N-K edge. Then
DNA stack file is formed by scaling normalized DNA
NEXAFS spectrum with the DNA peak height by
DNA quantity in each pixel of DNA image. DNA stack
file is subtracted from the image stack file. Since theresultant image stack file has no DNA component, thesimilar procedure can be applied to obtain a stack fileof other molecules one by one using possible specific
NEXAFS peak for each molecule.
Figure 1 shows chromosome image by STXM at
400 eV. The calculated DNA, histone and PC images

are shown in Fig. 2a. The discrete distribution of DNA
was observed, which possibly reflects band structure
in a chromosome. In Fig. 2b images of the same
molecules obtained by SVD method were shown forcomparison. The images are not so resolved comparedwith images by our method, and the molecular
contents were largely different (data not shown). SVD
method may not be suitable for applying straightly tobiological specimens with complex molecularcomposition.
Fig. 1. STXM imageof chromosomes of
CHO cells at 400 eV.

(a)

(b)

Fig. 2. RGB expression of molecular distribution
in chromosomes.
DNA, protein, PC are displayed with red, green andblue, respectively. (a) present method, (b) SVD[1] H. Ade et al., Science, 258 (1992) 972.
[2] X. Zhang et al., J. Struct. Biol., 116 (1996) 335.
[3] K. Shinohara, A. Ito, K. Kobayashi, In: X-Ray
Microscopy and Spectromicroscopy, Springer-Verlag,
Berlin/Heidelberg (1998), pp. III-157-III-161.
[4] A. Ito, T. Ohigashi, K. Shinohara, S. Tone, M.
Kado, Y. Inagaki and N. Kosugi, UVSOR Activity
Report 2015 43 (2016) 143.

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Chemical Bonding State Change of Phosphodiester Bonds in Plasmid DNA
T. Ejima1, T. Ohigashi2, M. Kado3 and S. Tone4IMRAM, Tohoku University, Sendai 980-8577, JapanUVSOR Synchrotron Facility, Institute for Molecular Science, Okazaki 444-8585, Japan
3
Takasaki Adv. Rad. Res. Inst., QST, Takasaki 370-1292, Japan
4
Sch. of Sci & Technol., Tokyo Denki University, Hatoyama 350-0394, Japan1

2

During the apoptotic process, phosphodiester bonds
in DNA are cleaved then DNA strand is broken [1].
When the chemical bonding state change of thephosphodiester bond can be observed according toprogress of the DNA cleavage, the DNA cleavage in acell-nucleus will be visualized by utilizing thechemical bonding state change of the phosphodiester
bond. In advance of the visualization of the DNA
cleavage, the P-L2,3 absorption edge structure ofplasmid DNA is investigated, and the results of theedge structures are compared with those of some
phosphate materials, such as di-nucleotide, trinucleotide, AMP, and cAMP.
Measured samples were a commercially available
plasmid DNA that was air-dried as it was on a 50nm
thick SiN membrane and that was cleaved with EcoRI
and air-dried on a same 50 nm thick SiN membrane.
X-ray absorption spectroscopy measurement wasmade in STXM at BL4U of UVSOR. Energy
resolution E/δE was >1000 and spatial resolution was
50 nm. Measured spectra are shown in Fig. 1.
Spectral shapes of measured P - L2,3 XAS spectra
can be reproduced by 6 peaks, which are indicated byalphabets designated in the figure. Energy positions
and peak intensities of corresponding peaks between
the spectra are similar to each other except for peak D.
Spectral structures of di-nucleotide, tri-nucleotide,
AMP, and cAMP show similar shape and arecomposed of 6 peaks at the same energy range. Whenthe spectral shapes are compared with that of KH2PO4[2], which spectral shape show the six peaks, which
spectral shape show the six peaks, the peaks A and B,
the peaks C and D, and the peaks E and F will be
originated from P3s+O2p, P3p+O2p, and P3d+O2p,
respectively.
Peak intensity of peak D will be increased as theresult of DNA-cleavage reaction. The comparison of
the spectral shapes between the measured spectra andthat of KH2PO4 suggest that the difference of the DNA
cleavage is associated with the P3p+O2p bonds, andthat state of DNA cleavage in cell-nucleus may be
visualized by the intensity difference of peak D.
This work was supported by Grants-in-Aid for
Scientific Research (JSPS Kakenhi, Grant Number:
16H03902 and 17K190210). Prof. K. Shinohara andProf. A. Ito are thanked for discussions. Y. Inagaki isthanked for assistance of the measurements.

Ⅲ- 5

Fig. 1. P-L2,3 XAS spectrum of plasmid DNA that airdried as it is, (a), and that cleaved with EcoRI and airdried, (b). Dots represent the measurement results,
blue curves are deconvolution results, and red cures,the summation of the peaks.
[1] S. Tone et al., Exp. Cell Res. 313 (2007) 3635.
[2] S. O. Kucheyev, et al., Phys. Rev. B 70 (2004)
245106.

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Investigation of Origin and Evolution of Extraterrestrial Materials by
Visualizing Molecular Structure and Construction of Analytical Schemewith Facility-crossing Collaboration of Hayabusa2 Retuned Samples
M. Uesugi1, M. Ito2, N. Tomioka2, K. Uesugi1, A. Yamaguchi3, N. Imae3, T. Ohigashi4,
N. Shirai5, Y. Karouji6, T. Yada6, M. Abe6 and Y. Inagaki4
2

Ⅲ- 5

1
Japan Synchrotron Radiation Research Institute (JASRI/SPring-8), Sayo 679-5198, Japan
Japan Agency for Marine-Earth Science and Technology (JAMSTEC), Nankoku 783-8502, Japan
3
National Institute of Polar Research (NIPR), Tachikawa 190-0014, Japan
4
UVSOR Synchrotron Facility, Institute for Molecular Science (IMS), Okazaki 444-8585, Japan
5
Tokyo Metropolitan University, Hachioji 192-0364, Japan
6
Japan Aerospace Exploration Agency (JAXA), Sagamihara 252-5210, Japan

Extraterrestrial sample curation center (ESCuC) of
Japan aerospace exploration agency (JAXA) organizeda special team for the development of techniques anddevices for the handling, transfer and analysis ofsamples returned by Hayabusa2 spacecraft, from 2015
under the agreements of collaboration. The special teamconstitutes of members of institutes those having stateof-the-art analytical instruments and experiences ofcuratorial works of precious natural samples, but
beyond the specialists of the extraterrestrial materials[1-2].
We started development of new analytical protocols
of extraterrestrial materials, which includes method ofsample transfer, sample separation and data sharing,
through the rehearsals of the curation works for the
initial description of Hayabusa2-returned samples using
extraterrestrial materials. Currently, 5 Antarctic
micrometeorites (AMMs) provided by national instituteof polar research (NIPR) were imaged by synchrotron
radiation computed tomography (SR-CT) and x-ray
diffraction (XRD) at SPring-8, and investigated by highresolution field emission scanning electron microscopy
and energy dispersive spectroscopy (FE-SEM-EDS)
system at institute for molecular sciences (IMS) (Fig. 1).
Through the series of non-destructive analysis, weselected Antarctic micrometeorites those having similar
characteristics of carbonaceous chondrites.
In this study, we formed thin sections of two AMMs
selected by the results of non-destructive analyses,
using focused ion beam (FIB) for the characterization oforganic materials by scanning transmitted x-ray
microscopy and x-ray absorption near edge structure
analysis (STXM-XANES) at BL4U. One section for a
fragment of AMM, TT006cT0009-1-1 was analyzed bynano-scale secondary ion mass spectrometry(NanoSIMS) before the STXM-XANES, and obtained
isotopic mapping images of the light elements such ashydrogen, carbon, nitrogen and oxygen by at Japanagency for marine-earth science and technology(JAMSTEC). After NanoSIMS analysis, the section
was fabricated by FIB again, to make ultrathin section
for the STXM-XANES analysis. Other sections wereprepared without NanoSIMS analysis.
Figure 2 shows STXM image of TT006cT0009-2-1
obtained using a NanoAnalysis linkage grid (Kochi

grid) developed in our activity. We could safely prepare
the sample in atmosphere shielding environment using
glovebox and sample loading system equipped at BL4U,
and investigated the ultrathin-section without paper tape.
Further result of our analyses will be reported bysubsequent papers (in prep).
After the STXM-XANES analysis, the ultrathinsections will be analyzed by transmission electron
microscopy (TEM), for high resolution analysis of
nano-scale textures and crystal structures inside the
AMMs.

Fig. 1. An energy dispersion spectroscopy image of
TT006cT0009-2. Red shows carbon distribution.

Fig. 2. A STXM-XANES image of a ultrathin-sectionof TT 006cT0009-2. Green portion shows regionenriched in graphene, and red portion shows regionenriched in carboxyl and poor in graphene.
[1] M. Uesugi et al. Abstract Haybusa symp. (2016)#1129[2] M. Uesugi et al. Abstract Haybusa symp. (2017)
#1129

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Probing the Penetration of Tofacitinib and Tofacitinib Citratein Inflamed Murine Skin by Soft X-Ray Spectromicroscopy
K. Yamamoto1, A. Klossek1, J. Berkemeyer1, T. Ohigashi2, F. Rancan3, R. Flesch1,
A. Vogt3, U. Blume-Peytavi3, M. Radbruch4, H. Pischon4, A. D. Gruber4, L. Mundhenk4,
N. Kosugi2 and E. Rühl1
1

Institute for Chemistry and Biochemistry, Freie Universität Berlin, Takustr. 3, 14195 Berlin, Germany
2
UVSOR Synchrotron Facility, Institute for Molecular Science, Okazaki 444-8585, Japan
3
Charité Universitätsmedizin, 10117 Berlin, Germany
4
Veterinary Pathology, Freie Universität Berlin, 14163 Berlin, Germany

The penetration of hydrophilic and hydrophobic
drugs into human skin is systematically investigatedby label-free detection employing soft X-ray
microscopy. The FDA-approved anti-inflammatory
drug tofacitinib citrate (C16H20N6O · C6H8O7,
molecular weight 504.49 g/mol), marketed under thename Xeljanz, is chosen as a hydrophilic drug, whichacts as a JAK inhibitor by interfering with the
JAKSTAT signaling pathway. It is a low molecularweight drug that is commonly used for oral drugdelivery for treating e.g. rheumatoid arthritis. Its use
for treating plaque psoriasis is currently underinvestigation [1]. We also included the free base of
tofacitinib (C16H20N6O, molecular weight 312.37
g/mol) as a hydrophobic complement to tofacitinib
citrate. This enables us to investigate the influence ofhydrophilicity on dermal drug penetration.
The experiments made use of excised murine skin,which was inflamed and exposed to tofacitinib citrate
and the free base obtained from LC Labs forpenetration times between 10 and 1000 min, similar toearlier work using dexamethasone as a drug [2-3].
Spectromicroscopy studies were performed at the
BL4U at UVSOR III using a scanning X-ray
microscope. The chemical selectivity for probing the
drugs in EPON-fixed skin was achieved by exciting
the samples selectively at the O 1s-edge in the preedge regime, i.e. at 528 eV (pre-edge) and 531.8 eV,
respectively, corresponding to the O 1s π* transitionof the drugs. Figure 1 shows the O 1s regime of fixedmurine skin, tofacitinib, and tofacitinib citrate.
Figure 1. This provides chemical selectivity forlabel-free probing of tofacitinib. Differential
absorption maps corresponding to the drug
distribution as a function of depth were derived similar
to our previous work [2-3]. Figure 2 shows the drug
distribution in the top skin layers, consisting of stratum
corneum (SC) and viable epidermis (VE). There are
distinct differences between the drug distributions
after 100 min penetration time. The hydrophilic drug
tofacitinib citrate is primarily found in the top layersof the stratum corneum and no drug is found in deeper
skin layers (see Fig. 2(a)). In contrast, the free base isfound in deeper layers of the stratum corneum and in
the viable epidermis (see Fig. 2(b)). There, the drug ispreferably found in the thin membranes of thekeratinocytes surrounding the cell nuclei. This
corresponds to the site of action of this drug. The
present results also indicate that topically appliedhydrophobic drugs can more efficiently penetrate the
top layers of skin than the hydrophilic analogue.

Fig. 1. X-ray absorption of tofacitinib citrate, the freebase, and fixed murine skin near the O 1s-edge.

Ⅲ- 5Fig. 2. Differential absorption of (a) tofacitinib citrateand (b) the free base of tofacitinib penetrating the top
skin layers of murine skin for 100 min, respectively.
High drug concentration is indicated by yellow color.
[1] V. Di Lernia et al., Drug Design Develop. Therap.
10 (2015) 533.
[2] K. Yamamoto et al., Anal. Chem. 87 (2015) 6173.
[3] K. Yamamoto et al., Eur. J. Pharm. Biopharm. 118(2017) 30.

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Chemical Mapping Individual Atmospheric Nanoparticles
X. Kong1*, M. Huttula1, J. J. Lin1, P. Corral Arroyo2, T. Ohigashi3,
N. Kosugi3, 4, Z. Wu5 and N. L. Prisle1

Nano and Molecular Systems Research Unit, University of Oulu, Oulu 90041, Finland
Laboratory of Environmental Chemistry, Paul Scherrer Institute, CH-5232 Villigen PSI, Switzerland
3
UVSOR Synchrotron Facility, Institute for Molecular Science, Okazaki 444-8585, Japan
4
School of Physical Sciences, The Graduate University for Advanced Studies, Okazaki 444-8585, Japan
5
State Key Joint Laboratory of Environmental Simulation and Pollution Control, College of Environmental
Sciences and Engineering, Peking University, 100871, China*
now at Department of Chemistry and Molecular Biology, University of Gothenburg, Gothenburg, Sweden
1

2

Ⅲ- 5

During the STXM beamtime, chemical maps weredrawn for two laboratory-generated aerosol samples
and one ambient urban aerosol sample.
The laboratory-generated aerosols were producedby the atomization of solutions of sodium n-decanoate
and sodium chloride. Particles of 150, 200, and 300
nm were generated from solutions with 0.5 and 0.8
mass fraction of sodium n-decanoate and particles of30, 50, 80 nm, and polydisperse samples weregenerated from solutions of 0.25 mass fraction of
sodium n-decanoate.
The urban aerosol samples were collected from
October 2017 to January 2018 in a winter campaign
jointly operated by Peking University and the
University of Gothenburg. Sampling was carried out
at the Peking University Atmosphere Environment
Monitoring Station on the roof of a six-floor buildingon the campus of Peking University located in thenorthwestern urban area of Beijing. A SKC 4-stage
cascade impactor was used to collect particles with
cut-off sizes of 250 and 500 nm that were impacted
onto Formvar films.
The urban aerosol samples were examined at theadsorption edges of carbon, nitrogen, sulfur, silicon,
chlorine, and potassium. Figure 1 shows the opticaldensity of a particle at the carbon and nitrogen edges.
At 284.5 eV, the C=C double bond is shown as thebright area [1], which indicates two soot components
located at the two poles of the particle. The spatial
resolution of the image is 30 nm. The detailed carbon
edge spectrum indicates that the soot particles are well
aged. In addition, the potassium L-edge indicates that
K+ ions are present and located in the area between thesoot particles. The absorption maxima of theammonium functional group are around 401 eV, andaccording to the panel (b) the NH4+ mainly stays
between the soot particles, like K+. From panel (c) it
seems that the NO3- is spread all over the particle.
Further quantitative analysis will be performed to
calculate the mixing ratio.
The carbon edge adsorption spectrum contains richinformation about the functional groups and theoxidation states. Figure 2 shows (a) the carbon spectraof aged soot particles at the carbon edge from 280 eV to
320 eV, where the spectra of different colors are takenfrom different places of the particle, but they are very
consistent except for the amplitudes that is correlated tothe particle thickness of the examined regions. Panel (b)shows the spectra of laboratory- generated soot coatedby α-pinene (in red) and naphthalene SOA (in black)from literature [2], where the featured peaks excellently
match the sampled particle shown in panel (a). The
STXM method at BL4U seems to be perfectly suited forthe study of both ambient and laboratory nanoparticles.

soot

NH4+

NO3-

Fig. 1. Optical density at various photon energies.
The bright areas indicate where the excited functionalgroups are located. (a) soot; (b) NH4+; (c) NO3-.

Fig. 2. (a) carbon K-edge adsorption spectra ofsampled urban particles; (b) STXM spectra oflaboratory-generated soot coated by α-pinene (red)and naphthalene SOA (black) [2]; (c) STXM image ofthe particles at 286.6 eV.
[1] R. C. Moffet, A.V. Tivanski and M.K. Gilles,“Fundamentals and Applications in Aerosol
Spectroscopy”(CRC Press; Boca Raton, Fl, 2011).
[2] J. C. Charnawskas et al., Faraday Discussions, 200(2017) 165.

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STXM Studies of Abnormal Ultrastructural Features in FINCA Disease
Nano and Molecular Systems research unit, P.O. Box 3000, 90014 University of Oulu, Finland
PEDEGO Research Unit and Medical Research Center Oulu, University of Oulu and Oulu University Hospital,
PO Box 5000, 90014 University of Oulu, Finland
3
UVSOR Synchrotron Facility, Institute for Molecular Science, Okazaki 444-8585, Japan
4
Biocenter Oulu, PO Box 5000, 90014 University of Oulu, Finland
1

2

E.-V. Immonen1, 2, T. Ohigashi3, I. Miinalainen4, M. Patanen1, R. Hinttala 2, 4,
J. Uusimaa2, 4, N. Kosugi3 and M. Huttula1

In this work, we have used scanning transmission Xray imaging technique at BL4U to study human tissue
samples exhibiting abnormal ultrastructural features
related to FINCA disease. Recently, a novel, fatal
cerebropulmonary disease in children was characterizedwith fibrosis, neuro- degeneration and cerebral
angiomatosis (FINCA disease) [1]. The disease course
is progressive with multiorgan manifestations, leading
to death before the age of two years. Studies on autopsy
samples revealed interstitial fibrosis and previously
undescribed granuloma-like lesions in the lungs, andhepatomegaly related to widespread microvacuolar
hepatocyte fatty degeneration and hepatocellularnecrosis. All patients had a combination of mutations in
the NHLRC2 gene encoding an NHL repeat containing
protein 2 that is ubiquitously present in various types oftissues, from animals to bacteria. Still, no publishedresults exist on the functional role of the NHLRC2.
In order to learn more about the pathomechanism
behind FINCA disease, multidisciplinary approaches
are called for. These, among others, include
morphological and chemical analysis of the subcellularfeatures in the liver autopsy samples taken from the
FINCA patients. However, conventional imaging
techniques, such as electron microscopy, possess severe
pitfalls when it comes to chemical imaging, requiring
the utilization of the STXM method. Thus, we have
performed the first proof-of-principle STXM studies on
subcellular structural characteristics related to storage
diseases.
The samples (approximately 150-200 nm thick) wereprepared at Biocenter Oulu, Finland using standard
methods in transmission electron microscopy (TEM),
and placed on Butvar film coated copper grids. In
addition, sections adjacent to ones selected for STXM
imaging were imaged using TEM, allowing thecomparison of these methods as shown in Fig. 1.
The measurements concentrated on C 1s edge, andwe have observed several abnormal ultrastructuralfeatures in various tissues of a FINCA patient. Figure
1 shows an example of a liver sample, where theseabnormal features include large vacuoles of unknown
origin (marked as U in Fig. 1). According to carbon
K-edge absorption profile, these vacuoles have achemical signature that differs from other organelles(Fig. 1C). Further studies will focus on verifying theorigin and identifying the accumulated compounds.

This can be useful in finding novel biomarkers infuture for example storage diseases.

Fig. 1. A) A TEM image showing organelles (lipid
vesicles L, their membrane MR, mitochondria M, andan unknown structure U, within a hepatocyte. B) An
STXM image of the same region with hν = 287 eV.
Subcellular features used for collecting region specificabsorption spectra are marked with colors. C)
Absorption spectra from regions in B. The arrows
indicate the most prominent spectral differences. D)
STXM image stack from an energy range 285-286 eV(1st arrow in C). E) Energy range 286-288 eV (2ndarrow in C). F) 288-297 (3rd arrow in C).
[1] J. Uusimaa, R. Kaarteenaho, T. Paakkola, H.
Tuominen, M. K. Karjalainen, J. Nadaf, T. Varilo, M.
Uusi-Mäkelä, M. Suo-Palosaari, I. Pietilä, A. E.
Hiltunen, L. Ruddock, H. Alanen, E. Biterova, I.
Miinalainen, A. Salminen, R. Soininen, A. Manninen,
R. Sormunen, M. Kaakinen, R. Vuolteenaho, R. Herva,
P. Vieira, T. Dunder, H. Kokkonen, J. S. Moilanen, H.
Rantala, L. M. Nogee, J. Majewski, M. Rämet, M.
Hallman, R. Hinttala, Acta Neuropathol. (2018). In
press. https://doi.org/10.1007/s00401-018-1817-z

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