Ⅲ-5

Life, Earth andPlanetary Sciences

BL1UⅢ -5. Life, Earth and Planetary Sciences

BL1U

Optical Activity Emergence in Amino-Acid Films by Vacuum-Ultraviolet
Circularly-Polarized Light Irradiation (III) - Summary of Experiments J. Takahashi1, T. Sakamoto1, Y. Izumi2, K. Matsuo2, M. Fujimoto3, M. Katoh2,3,
Y. Kebukawa1 and K. Kobayashi1
Faculty 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

One of the most attractive hypotheses for the originof homochirality in terrestrial biomolecules (L-aminoacid and D-sugar dominant) in the context ofastrobiology is “Cosmic Scenario” [1,2]. Several
ground experiments to validate the scenario have been
investigated using circularly polarized light (CPL)from high-energy particle accelerators. We havealready reported optical activity emergence in solid
films of racemic amino acids by CPL irradiation of 215
nm in wavelength from free electron laser (FEL) of
UVSOR-II [3].
Circular dichroism (CD) spectroscopy can detect
optical activity emergence with a high accuracy
because CD spectra sensitively reflects the stericstructures of chiral molecules. The theoretical
calculation of CD spectrum of L-alanine molecule hasrevealed that the absorption bands of carboxyl andamino groups are derived from electronic transitions(n-π*, π-π*, n-σ*) corresponding to wavelengths
below 230 nm (Fig.1) [4]. It is suggested that opticalactivity emergence by asymmetric photochemical
reactions depends on the photon energy of CPL.
We carried out irradiation experiments by using
CPL with different wavelengths to investigate thephoton energy dependence of photochemical chiralreactions. We formed thin solid films of racemic
mixture of alanine on quartz substrates from crystal
powders of DL-alanine by using a thermal-crucible
vacuum-evaporation system. Sublimation temperature
was controlled in the range of 150~200°C and pressureof the vacuum chamber was approximately 5x10-2 Pathroughout the evaporation process. The CD spectra ofthin solid films just after the deposition were measured
from 260 to 160 nm in wavelength using a vacuumultraviolet CD beam line BL-12 of HiSOR andconfirmed to be mostly zero CD before the irradiation,
showing that the spurious CD due to the contaminationof film surface were negligible.
The thin solid films of DL-alanine were irradiatedwith CPL in different wavelengths using the undulator
beamline BL1U of UVSOR-III. In case of CPL
irradiation in shorter wavelengths than 200 nm, thesamples were set in a vacuum sample chamber
preventing attenuation by air absorption. On the beam
entrance side of the vacuum sample chamber, a gate
valve with an MgF2 vacuum sealing window was
mounted. The irradiated CPL wavelengths were 180
and 155 nm corresponding to absorption bands ofalanine molecule. The irradiated photon energy dose
was measured with photoelectron current of a silicon
photodiode (International Radiation Detectors, Inc.)
settled at the sample position.
CD spectra of the CPL irradiated films were
measured at BL-12 of HiSOR to clarify the opticalactivity emergence by CPL irradiation. In order todelete the effects of linear dichroism (LD) and/or
linear birefringence (LB) components, the CD spectra
were calibrated by measuring the dependence on
sample rotation angle (0, 45, 90, and 135 degrees). As
the results, the dependence of the CD spectra on theirradiated wavelength and also on the polarization(left- or right-handed) of CPL have been clearlyobserved (Fig.1) [5,6]. Detailed analysis of CD spectra
is in progress to clarify full mechanism of the opticalactivity emergence, which potentially has relevance tothe origin of terrestrial bioorganic homochirality
stimulated by CPL.

Ⅲ- 5

Fig. 1. CD spectra of DL-alanine film after left (L)or right (R)-handed CPL irradiation.

[1]W. A. Bonner, Orig. Life Evol. Biosph. 21 (1991) 407.
[2] J. Takahashi et al., Symmetry 11 (2019) 919.
[3] J. Takahashi et al., Int. J. Mol. Sci. 10 (2009) 3044.
[4]F. Kaneko et al., J. Phys. Soc. Jpn. 78 (2009) 013001.
[5] J. Takahashi et al., UVSOR Activity Report 2018 46(2019) 156.
[6] J. Takahashi et al., Proc. The 23rd Hiroshima Int.
Symp. on Synchrotron Radiation (2019) 55.

143
Ⅲ05- 1 -Ⅲ -5. Life,BL1U
Earth and Planetary Sciences

BL1U

Laser Compton Scattering Gamma-ray Generation for Nonlinear Effectin QED
T. Hayakawa1,2, K. Kawase1, T Shizuma1, J. K. Koga3, H. Zen4, T. Kii4, H. Ohgaki4,
M. Fujimoto5 and M. Katoh5

Tokai Quantum Beam Science Research Center, National Institutes for Quantum and Radiological Science andApplications, Tokai 319-1106, Japan
2
National Astronomical Observatory of Japan, Mitaka 181-8588, Japan
3
Kansai Photon Science Institute, National Institutes for Quantum and Radiological Science and Applications,
Kizugawa 619-0215, Japan
4
Institute of Advanced Energy, Kyoto University, Uji 611-0011, Japan
5
Institute for Molecular Science, National Institutes of Natural Sciences, Okazaki 444-8585, Japan

1

Ⅲ- 5

QED predicted unresolved nonlinear effects such as
photon-photon interactions. However, because theircross sections are extremely small, the interactions
have not been well experimentally measured [1,2].
Delbrück scattering, in which a gamma-ray interactingwith a Coulomb field creates an electron-position pair,which subsequently annihilates to generate a gammaray whose energy is almost identical with the incident
gamma-ray, is one of important phenomena to study
nonlinear effects by QED and vacuum polarization.
Koga and Hayakawa [3] have presented that it ispossible to measure selectively the amplitude ofDelbrück scattering using linearly polarized gammaray beams. Furthermore, if one uses a linearlypolarized beam with energies lower than 1.022 MeV,
which is the threshold of the pair creation, it is possible
to measure only the virtual process of Delbrück
Scattering, namely vacuum polarization.
For such a purpose, we have developed a laser
Compton scattering (LCS) gamma-ray beam with a
CO2 laser having a wavelength of 10.6 μm at the BL1U
beamline in UVSOR-III, where the energy of theelectron beam stored in top-up mode is approximately
750 MeV. We have demonstrated the 1-MeV LCS
gamma-ray beam generation using a randomly
polarized CO2 laser [4].
We newly installed a high power linearly polarized
CO2 laser in order to generate linearly polarized LCS
gamma-ray beam. The maximum power of the laser is
130 W. We measured the energy spectra of thegenerated LCS gamma-ray beam using 3.5”  4”
LaBr3(Ce) scintillation detector, and the energy
spectra of the photon scattered from a tin or tungsten
target with a diameter of 5 mm and a length of 50 mm
using two high-purity Ge detectors (see Fig. 1). Figure
2 shows a typical measured energy spectrum. The
Compton scattering from the target is clearly observedin the energy region of 400-600 keV. The gamma-ray
from natural backgrounds are also observed.

Fig. 1. Photo of the two high-purity Ge detectors at
the BL1U beamline.

Fig. 2. Typical spectrum of scattered photons fromthe target, measured by the Ge detector.
[1] ATLAS Collaboration, Nat. Phys. 13 (2017) 852.
[2] T. Inada et al. Phys. Lett. B 732 (2014) 356 .
[3] J. K. Koga and T. Hayakawa, Phys. Rev. Lett. 118(2017) 204801.
[4] H. Zen et al., J. Phys.: Conf. Ser. 1067 (2018) 092003.

144
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Ⅲ -5. Life, Earth and Planetary Sciences

BL3U

X-ray Absorption Spectra of Lipid Molecules in Bilayer Membranes
Measured in Aqueous Solution
R. Tero1, W.-Z. Goh1 and M. Nagasaka2

Toyohashi University of Technology, Toyohashi, 441-8580, Japan
2
Institute for Molecular Science, Okazaki 444-8585, Japan1

The lipid bilayer is a self-assembled structure ofamphiphilic lipid molecules, and is the fundamental
structure of biomembranes such as cell membranes.
Internal structures of lipid bilayers, e.g. two dimensional
domains and hydrophobic thickness, and physicalproperties affect the transportation of materials,
information and energy through the biomembranes. All
these physiological reactions proceeds in the presence ofions. Ions in the aqueous solution significantly influence
to these structures and properties of lipid bilayers.
Phosphatidylcholine (PC) (Fig. 1 (a)) is the major
component of eukaryotic cells. Cations bind to thephosphate and carbonyl groups of PC. However, affinityof cations to PC, and also other lipids, are still
controversy especially in the fields of theoreticalsimulations [1]. We aim to determine the binding affinityof cations to lipids in aqueous solutions experimentally,by means of X-ray absorption spectroscopy (XAS) [2,3].
We prepared vesicle suspensions of dioleoyl-PC(DOPC) (Fig. 1 (a)), dioctadecenyl-PC (dietherPC) (Fig.
1 (b)), and digalactosyldiacylglycerol (DGDG) (Fig. 1(c)) in a buffer solution. We introduced the suspension
into the XAS flow cell consisting of Si3N4 membranes,
to form planar lipid bilayers on the Si3N4 surface
through the process of vesicle fusion method [3].
Fluorescence microscope imaging and fluorescencerecovery after photobleaching method showed that
DOPC, dietherPC, and DGDG bilayers fully covered
the Si3N4 membrane surface.
Figure 2 shows O K-edge spectra of the planar lipid
bilayers, after subtracted the spectrum of the Si3N4
membrane before the lipid bilayer formation. DOPC,
which has phosphate and carbonyl groups (Fig. 1 (a)),
showed at least two components at 531.06 and 531.94
eV. To identify these components, we measured XAS
spectra of dietherPC and DGDG, which have only
phosphate group and carbonyl group, respectively (Fig.
2). DietherPC bilayer showed two components at
531.34 and 532.22 eV. We attributed them to 1s→π*transitions of P=O and P-O-Na, respectively, on thebasis of inner shell calculation. DGDG bilayer had
dominant peak at 532.02 eV. Therefore, the DOPC
component at 531.06 eV is attributed to P=O, and thatat 531.94 eV to overlapping P-O-Na and C=O.
In conclusion, we obtained XAS spectra of lipid
bilayers in an aqueous solution, and identified thecomponents in the spectra. We expect these XAS
spectrum components provide information of the effectsof cation species and concentration on lipid bilayers.

Fig. 1. (a) Dioleoylphosphatidylcholine,(b) dioctadecenylphosphatidylcholine, and(c) digalactosyldiacylglycerol.

Ⅲ- 5

Fig. 2. O K-edge spectra of (a) DOPC, (b) dietherPC,and (c) DGDG bilayers.
[1] T. B. Woolf, Biophys. J. 104 (2013) 746.
[2] M. Nagasaka, H. Yuzawa, T. Horigome and N.
Kosugi, J. Electron Spectros. Relat. Phenomena 224(2018) 93.
[3] R. Tero, S. Nakamura, Y. Sano and M. Nagasaka,
UVSOR Activity Report 2018 46 (2019) 158.

145
Ⅲ05- 3 -Ⅲ -5. Life,BL4U
Earth and Planetary Sciences

BL4U

P-L2,3 Absorption Spectra of Phosphates
in Plasmid DNA and Adenosine Compounds
2

Ⅲ- 5

T. Ejima1, T. Ohigashi2 and S. Tone3

IMRAM, Tohoku University, Sendai 980-8577, JapanUVSOR Synchrotron Facility, Institute for Molecular Science, Okazaki 444-8585, Japan
3
Sch. of Sci & Technol., Tokyo Denki University, Hatoyama 350-0394, Japan1

DNA is composed of simpler monomeric units called
nucleotides. The nucleotides are joined to one anotherin a chain by covalent bonds between the sugar of one
nucleotide and the phosphate of the next, resulting in analternating sugar-phosphate backbone. The phosphate
PO43– forms ideally a tetrahedral structure in which fouroxygen atoms coordinate equidistantly from a center P
atom.
The NEXAFS spectrum of the P-K absorption edge
in DNA is similar to those of aqueous phosphate(H2PO4), biological solids, and animal manures [1,2].
This similarity will be originated from the stable
tetrahedral-structure of the phosphate PO43–. On theother hand, the spectral differences among thephosphate compounds at the P-L2,3 absorption edges are
remarkable even though the energy positions of the PL2 and P-L3 edges are close to each other [3]. Therefore,
it seems natural to focus on P-L2,3 absorption edges
when shapes and positions of DNA are observed using
an X-ray microscope, which is expected to have a highspatial resolution because of the shortness of theobservation wavelength [4]. In this study, we focused on
the P-L2,3 absorption edges (~135 eV) and acquired
absorption spectra for the visualization of DNA using
the P absorption edges.
Two Adenosine compounds and two DNA were
measured: adenosine phosphate (AMP), adenylyl- (5'3')-adenylyl- (5'-3')-adenosine (AAA), plasmid DNA
with a circular structure, and plasmid DNA with afibrous structure.
The obtained spectra are shown in Figs. 1 and 2.
Peak structures are indicated from A to G in the figuresand each peak exists at almost the same photon energy
positions. The difference among the spectra appears
mainly at the peak E.
The density of states of AMP and AAA were
calculated to understand the origin of the spectral
difference, and the results showed that the P 3d statesof AMP and AAA are mainly hybridized with P 3s
states and located at around the energy position of peak
E. Hence the similarity of the spectral structures
between the figures may suggest that the difference ofthe peak E will be interpreted as the P 3d states
difference between the DNAs. Further calculationreflecting the structural difference between the DNAs
is necessary for the interpretation of the spectral
difference in DNAs.

Fig. 1. P-L2,3 XAS spectrum of AMP and AAA. Dots
represent the measurement results, solid and broken
curves are smoothed data.

Fig. 2. P-L2,3 XAS spectrum of plasmid DNA that airdried as it is (“uncut”), and that cleaved with EcoRI
and air-dried (“EcoRI”).
[1] A. Ito, K. Shinohara, Y. Mizukami, H. Nakano, K.
Yada, T. Uehara and T. Honda, J. Synchrotron Rad. 5(1998) 1099.
[2] A. L. Shober, D. L. Hesterberg, J. T. Sims and S.
Gardner, J. Environ. Qual. 35 (2006) 1983.
[3] J. Kruse, P. Leinweber, K.-U. Eckhardt, F. Godlinski,
Y. Huc and L. Zuinc, J. Synchrotron Rad. 16 (2009) 247.
[4] D. Attwood and A. Sakdinawat, "X-rays and Extreme
Ultraviolet Radiation" (Cambridge Univ. Press, 2016,
Cambridge, UK) Chap. 11.
[5] W. R. Hunter, "Vacuum Ultraviolet Spectroscopy"(ed. by J. A. Samson and D. L. Ederer, Academic Press,
2000, San Diego, USA) Chap. 11.

146
Ⅲ05- 4 -BL4U

Ⅲ -5. Life, Earth and Planetary Sciences

BL4U

STXM/XANES Analysis of a Carbonaceous Chondrite Lithology
in the Almahata Sitta Meteorite
Y. Kebukawa1, T. Ohigashi2 and M. E. Zolensky3

Faculty of Engineering, Yokohama National University, Yokohama 240-8501, JapanUVSOR Synchrotron Facility, Institute for Molecular Science, Okazaki 444-8585, Japan
3
ARES, NASA Johnson Space Center, Houston, TX 77058, USA
1

2

The Almahata Sitta (AhS) meteorite is a brecciated,
polymict ureilite that has originated from the nearearth asteroid 2008 TC3 which is classified as an Ftype asteroid (a subtype of C complex asteroids) [1].
Carbonaceous chondritic lithologies AhS 671 and AhS
91/91A are known to consist of aqueously alteredmineralogy such as phyllosilicates, bruennerite,
dolomite, magnetite, fayalite, ilmenite, phosphates,
pyrrhotite, and pentlandite, and have similarities to
CI1 chondrites, but shows evidence of heterogeneous
thermal metamorphism [2]. The textures of AhS 91A
and AhS 671 support formation of 2008 TC3 in aregolith environment, and could represent a volume ofregolith formed when a carbonaceous chondrite-likebody impacted into ureilitic body [2]. Althoughureilites usually contain no or little organic matter, it ishighly possible that the CI1 lithologies of AhS containabundant organics as in the case of CI1 chondrites (e.g.,
Orgueil and Ivuna). To understand the nature oforganic matter in CI1 lithologies, and similarity andrelationships to primitive carbonaceous chondrites, weanalyzed AhS 91A using the scanning transmission Xray microscope (STXM) at BL4U.
A 100 nm-thick section from AhS 91A was preparedusing a focused ion beam equipment at NASA-JSC. C,
N, O, Fe-X-ray absorption near-edge structure (XANES)
spectra of the sections were obtained using the STXM
on BL4U at the UVSOR.
First, we obtained a C-map of the AhS 91A FIB section
to locate organic matter. Carbonaceous compounds were
distributed partially in the FIB section in sub-micrometerscale (Fig. 1 (a)). Then, “energy stacks” were obtained at
C-rich areas. Selected C-XANES spectra generated fromthe energy stacks are shown in Fig. 1 (b). The C-XANES
spectra of the organic-rich regions show large peaks at
284.8 eV due to aromatic carbon, smaller peaks at 286.3
eV and 288.4 eV due to C=O and C(=O)O groups,
respectively.
These futures are often observed in aqueously alteredcarbonaceous chondrites (CI, CM, and CR chondrites),with some differences in the intensity ratios of thesepeaks [e.g., 3]. Some areas are rich in carbonate which
is characterized by a large peak at 290.4 eV. There is no1s-σ* exciton peak at 291.7 eV of graphene structuresthat is characteristic of thermally metamorphosedchondrites [4]. The molecular structures of organics aswell as carbonates, support that the AhS 91A isoriginated from a aqueously altered carbonaceouschondrite-like parent body [2]. Although someheterogeneous thermal metamorphism are observed in

the AhS 91A, the analyzed region of the AhS 91A has
not experienced significant heating events.

Ⅲ- 5

Fig. 1. (a) C-map and (b) C-XANES spectra of theselected regions in the FIB section of AhS 91A.

[1] P. Jenniskens et al., Nature 458 (2009) 485.
[2] C. A. Goodrich et al., Meteoritics & Planet. Sci. 54(2019) 2769.
[3] C. Le Guillou et al., Geochim. Cosmochim. Acta
131 (2014) 368.
[4] G. D. Cody et al., Earth Planet. Sci. Lett. 272 (2008)
446.

147
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Earth and Planetary Sciences

BL4U

Trial of Molecular Mapping for Thin Sections of Isolated Mammalian
Nuclei Embedded in Resin Using STXM
A. Ito1, K. Shinohara2, A. Matsuura3, S. Toné4, H. Yuzawa5 and T. Ohigashi5,6School 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
UVSOR Synchrotron Facility, Institute for Molecular Science, Okazaki 444-8585, Japan
6
School of Physical Sciences, The Graduate University for Advanced Studies, SOKENDAI, Okazaki 444-8585, Japan
1

2

Ⅲ- 5

Spectromicroscopy using scanning transmission Xray microscope (STXM) has been widely accepted as a
useful tool to visualize molecular distribution in
specimens. Using STXM installed at BL4U of UVSOR,
we have been developing an image processing
procedure for the quantitative mapping of biomolecules,
and successfully applied to the distribution of nucleic
acids (DNA and RNA) and proteins such as histone andbovine serum albumin (BSA) in biological specimens
using combined NEXAFS at the C, N and O-K
absorption edges [1-3]. At present, it is difficult toobtain reliable results for thick specimens such as
nucleus area in a whole mammalian cell or an isolated
nucleus, due to insufficient transmitted photons throughthe specimens [3].
Sectioning of a specimen with a thickness as thin as
the submicron level is one possible way to cope withsuch problem. Usually thin sections are made with amicrotome for resin embedded specimens. In thepresent study, we tried to study the thin section of
nucleus embedded in organic resin.
Isolated nuclei from human HeLa S3 cells wereembedded in Quetol-812 epoxy resin with curing agents,
DDSA (dodecenyl succinic anhydride) and MNA(methyl nadic anhydride), and a curing promoter DMP30 (2,4,6-tri(dimethylaminomethyl) phenol) (Nissin
EM Co., Ltd., Japan). The section of the resin was madeto a thickness of 0.5 m, and attached on an address
mesh (HF-15, Nissin EM) supported with collodion
film.
We applied our method to calculate the distributionsof nucleic acids and proteins [3] to the observed data forthe section specimens. Figure 1 shows the X-ray imageof the isolated nucleus at 294 eV (panel a), andcomposite image of DNA, RNA and histone displayedwith different colors (panel b). Contrary to our
expectation to be DNA, the main component in the
nucleus was found to be RNA. The cause was suspected
to be on an analytical process probably due to that the
major contribution to the absorbed data was resin.
To improve this situation, we made assumption thatimages for residual data (spect3) are mainly composedof resin. We generate energy stack file for resin from thespect3 and subtracted it from the original data at the firststep of analysis. Our calculation procedure was applied
to the thus-obtained stack file. The results are shown in

Figure 2. Three colors, red, green and blue, are given toDNA, RNA and histone for panel a, DNA, RNA andBSA for panel b, and DNA, histone and BSA for panelc, respectively. DNA was found to the major component
in a nucleus, which was in accord with our previous
observations for a chromosome [1], and a whole cell [2].
Further analysis is planned to apply to isolated nuclei
which undergo apoptosis.
We thank Prof. K. Tohya of Kansai University of
Health Sciences for his kind help with making thinsections.
(a)(b)

Fig. 1. Isolated nucleus of human HeLa S3 cell.
(a) X-ray image at 294 eV, (b) molecular distribution
in the nucleus. Red: DNA, Green: RNA, Blue: histone.

(a)

(b)

(c)

Fig. 2. Improved molecular distribution in the isolated
nucleus. (a) DNA (Red), RNA (Green), histone (Blue),(b) DNA (R), RNA (G), BSA (B), (c) DNA (R), histone(G), BSA (B)[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.

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Ⅲ -5. Life, Earth and Planetary Sciences

BL4U

In situ Analysis for Structure Change of Extraterrestrial Organic Materials
During Heating
M. Uesugi1, M. Ito2, K. Tomioka2, Y. Kodama3, T. Ohigashi 4,5, H. Yuzawa4, K. Uesugi1,
A. Yamaguchi6, N. Imae6, Y. Karouji7, N. Shirai8, T. Yada9 and M. Abe91
Japan synchrotron radiation research institute (JASRI/SPring-8), Say-cho 679-5198, Japan
Kochi Institute for Core Sample Research, Japan Agency for Marine-Earth Science Technology (JAMSTEC),
Nankoku 783-8502, Japan
3
Marine Works Japan Ltd., Yokosuka 237-0063, Japan
4
UVSOR Synchrotron Facility, Institute for Molecular Science, Okazaki 444-8585, Japan
5
School of Physical Sciences, The Graduate University for Advanced Studies, SOKENDAI, Okazaki 444-8585, Japan
6
Antarctic Meteorite Research Center, National Institute of Polar Research, Tachikawa 190-8518, Japan
7
JAXA Space Exploration Center, Japan Aerospace Exploration Agency (JAXA), Sagamihara 252-5210, Japan
8
Department of Chemistry, Graduate School of Science, Tokyo Metropolitan University, Hachioji 192-0397, Japan
9
Institute of Space and Astronautical Science, Japan Aerospace Exploration Agency (JAXA),
Sagamihara 252-5210, Japan
2

Organic compounds in extraterrestrial materials arethought to be one of the important components for thebuilding block of the Earth, especially for the origin oflife. Those organic materials would be transported to the
Earth mostly by micrometeorites, because previous
studies suggested that the flux of micrometeorites
falling to the Earth is much larger than that of meteorites(5.38 x 104 kg/y for meteorites and 4 x 107 kg/yr for
micrometeorites) [1,2]. In addition, micrometeorites
show distinct features that their chemical compositions
are rather similar to carbonaceous chondrites than any
other meteorites [3]. It is well known that mostmicrometeorites are heated up to 2000 ºC during theatmospheric entry, due to atmospheric drag heating.
During such intense heating under the oxidized
condition, organic compounds would be modified
and/or destructed. It is important to evaluate the effectof heating of organic compounds during theatmospheric entry, to investigate origin and evolution ofterrestrial organics.
We found that a number of organic nano-globules
survived in the Antarctic micrometeorites (AMMs)
larger than 150 μm. These AMMs would haveexperienced intense heating during atmospheric entry
due to large moments of inertia, in previous studies [4].
Numerical calculations showed that such AMMs
would have heated higher than 1000 ºC, and also
showed that ram pressure during the heating is almost
equivalent to 1 atm.
In order to investigate the possibility of survival oforganic matters during heating, we conducted heating
experiments of carbonaceous meteorites at 1 atm with
600 ºC in atmosphere and 400 ºC in N2 environment,
and evaluate the effect on the organic materials using
scanning transmission x-ray microscopy and near edge
x-ray absorption fine structure (STXM-NEXAFS)
analysis. The results of our experiments showed thatorganic materials were easily destructed anddisappeared in oxidized condition, while organics
became graphitic and did not disappear during heatingin N2 environment.

The results indicate that organic materials in
meteorites would not be destructed by heating in theinterplanetary space, because there are less oxygen andwater vapor. The organic materials would survive fromthe heating by irradiation of sunlight, during thetransportation from their parent body to the Earth. On
the other hand, the organic materials would be easily
damaged during the atmospheric entry because ofhighly oxidized atmosphere.
Survival of organic materials in AMMs suggeststhat incident velocities of the precursor meteorites arequite low, less around 1 km/s, and/or, their original size
is much larger than 1 cm and they would be destructed
into smaller pieces during the atmospheric entry. Thenunheated materials inside the meteorites can form
unheated AMMs.

Ⅲ- 5[1] Yada T. et al., Earth Planet Space 56 (2004) 67.
[2] Zolensky M. et al., Flux of extraterrestrial materials,
in Meteorites and the early solar system II (2006) 869.
[3] Imae N. et al., Geochim. Cosmochim. Acta 100(2013) 116.
[4] Uesugi et al., Preparation for the analysis of
Hayabusa2 returned samples JpGU meeting, abstract(2019).

149
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BL4U

The Behavior of Sulfur in the Past Martian Surface Water Based on
STXM-XANES Analysis of Nakhlite Y 000593
H. Suga1,2, K. Suzuki2, T. Ohigashi3,4, H. Yuzawa3,4, Y. Takeichi5, A. Yamaguchi6, T. Usui7,
Y. Takahashi2 and M. Miyahara8
Japan Synchrotron Radiation Research Institute (JASRI/SPring-8), Sayo 679-5198, Japan
2
The University of Tokyo, Tokyo 113-0033, Japan
3
UVSOR Synchrotron Facility, Institute for Molecular Science, Okazaki 444-8585, Japan
4
The Graduate University for Advanced Studies, SOKENDAI, Okazaki 444-8585, Japan
5
High-Energy Accelerator Research Organization (KEK), Tsukuba 305-0801, Japan
6
National Institute of Polar Research, Tachikawa 190-8518, Japan
7
Japan Aerospace Exploration Agency (JAXA), Sagamihara 252-5210, Japan
8
Hiroshima University, Higashi-Hiroshima 739-8526, Japan

1

Ⅲ- 5

Nakhlites are intriguing Martian meteorites because
they have various secondary minerals formed bywater-rock reactions on Mars [1]. The species,
assemblages, and chemical compositions of thesecondary minerals record the physicochemical
properties such as temperature and pH-Eh when thereactions occurred. Hence, the detailed understandingof the secondary minerals allows us to give a new
constraint on the physicochemical evolution of thesurface water on Mars.
The formation processes of the secondary minerals
in nakhlites are enigmatic because carbonate andsulfate minerals coexist in some nakhlites [1-3]: theformer precipitates under reductive alkaline condition,
the latter under oxidative acidic condition. We
examined the sulfide and sulfate minerals in the
petrologic thin section of nakhlite Y 000593 (#58-1
stored in NIPR) by FE-SEM-EDS/FE-EPMA at the
University of Tokyo, μ-XRF-XAFS at BL-15A of
Photon Factory, and STXM-XANES at BL4U of
UVSOR to elucidate the formation process of thesulfate minerals.
Y 000593 consisted mainly of olivine phenocryst
and mesostasis (clinopyroxene, plagioclase, and FeS).
Elemental mapping by EPMA showed that a trace
amount of sulfur (S) distributed in the carbonate
minerals occurring in the fractures of the olivine grains.
μ-XRF-XAFS analysis detected the existence of SO4
in the carbonate minerals. The abundance of S
decreased from the rim to the center of the olivine
grains. As for the origin of the SO4 (or sulfate
minerals), the only one S-containing mineral in Y
000593 FeS is the candidate.
FE-SEM-EPMA analysis revealed that most of the
FeS minerals in the mesostasis of Y 000593 was partly
altered. Hence, the oxidation state of the S in the FeS
minerals of the mesostasis was measured by μ-XRFXAFS. The altered portions of the FeS minerals weremainly composed of FeSO4.
We tried to clarify the dissolution path of SO42- fromthe FeS minerals to the surrounding minerals (finally,
into the fractures of the olivine grains) by FIB-STXM.
We made the ultrathin section including the partly

altered FeS minerals by FIB for the STXM analysis.
STXM Fe- and S L-edge XANES analysis revealed
that the sulfates dissolved from the FeS minerals into
the surrounding minerals (Fig. 1).
These results indicate that the origin of SO4 in the
carbonate minerals is the SO42- dissolved from the FeS
minerals. The chemical reaction formula could be
expressed as follows (as a simplified model):FeS + 5/2H2O + 3/2O2 + 1/3K+ →1/3KFe3(SO4)2(OH)6 + H+ + 1/3SO42FeS is easily oxidized by liquid water and becomes
sulfate mineral jarosite [KFe3(SO4)2(OH)6)]. Also, thisreaction makes sulfate-bearing acid fluid, which
decreases the pH. The SO42-(and/or jarosite) formed bythis reaction should be also the origin of the SO4
preserved in the carbonate minerals. Carbonate can beeasily dissolved by acid fluid. Therefore, we expectthat the sulfate minerals in Y 000593 have occurred
before the formation of the carbonate minerals as avein filling precursor.
(a)

(b)

5 μm

Fig. 1. FIB section scanned by STXM (white dashedline). (a) The red and dark green portions are FeS
mineral and plagioclase, respectively. (b) The blue andyellow portions are FeS and FeSO4, respectively. Fe3+
and SO42- dissolved from FeS mineral move into thefractures of plagioclase.

[1] L. M. White et al., Astrobiology 14 (2014) 170.
[2] T. Noguchi et al., JGR 144 (2009) E10004.
[3] T. Tomkinson et al., Nat. Commun. 4 (2013) 2662.

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Control of Drug Dermal Penetration: Quantitative Probing of Rapamycin
G. Germer1, T. Ohigashi2, H. Yuzawa2, R. Flesch1, F. Rancan3, A. Vogt3 and E. Rühl1
Physical Chemistry, Freie Universität Berlin, Arnimallee 22, 14195 Berlin, Germany
UVSOR Synchrotron Facility, Institute for Molecular Science, Okazaki 444-8585, Japan
3
Charité-Universitätsmedizin, 10117 Berlin, Germany
1

2

Progress in quantitative probing of topical drugdelivery is reported, which goes beyond our previous
work performed at UVSOR III [1,2]. Briefly,
hyperspectral imaging of dermal drug penetration isinvestigated using scanning transmission X-ray
microscopy (STXM). The experiments were performedat the BL4U beamline of UVSOR III. They mostly relyon scanning of the photon energy in the O 1s regime (520
– 560 eV), allowing for the acquisition of stacks ofimages by concentrating on the stratum corneum, the top
horny layer of skin. High spatial resolution,corresponding to pixel sizes of 50 nm, allows us togenerate in this heterogeneous target reliably reference
spectra of the crucial species contained in fixed humanskin, especially the topically applied drug rapamycin(C51H79NO13, M=914.13 g/mol), as well as identificationof different skin regions, i.e. corneocytes, lipid layers
between the corneocytes, and the fixation medium EPON.
Data reduction is performed by using a linear
combination of these site-dependently varying
contributions, similar to previous work [3]. This yields
the local composition of each component. The absoluteabsorption cross section of the drug is used to determine
quantitatively its local concentration and the relation to
the dermatologically relevant skin treatment conditions.
Characteristic results are summarized in Fig. 1, which
are part of systematic studies. Figure 1 (a) shows thestratum corneum of untreated skin, whereas Fig. 1 (b)shows skin that has been treated for 24 h by 0.1%
rapamycin in petroleum jelly. The optical density (OD)
recorded at 532 eV is shown in the maps labeled A (topplots in Figs. 1 (a), (b) ), clearly indicating the stratified
structure of the stratum corneum. The data analysis
shown in B clearly distinguishes corneocytes (Cor)(white) from lipids between the corneocytes (Lip) (black),
and the viable epidermis (VE) (grey). The maps labeled
C show the drug distribution with a clear enhancement, if
rapamycin is topically applied (see Fig. 1 (b) ). In D thedrug concentration is integrated over the maps shown in
C as a function of depth, yielding absolute concentrations
given in fmol/μm3, corresponding to a local maximum indrug concentration of 270 fg/μm3. The reference skinshown in Fig. 1 (a) indicates that no drug is found, but theerror bars are sizable due to the data reduction procedure.
The sample containing the topically applied drug inpetroleum jelly indicates that there are distinct variations
in local drug concentration due to the stratified structureof the stratum corneum, where local minima occur in thelipid matrix. This is unexpected for non-polar drugs, suchas rapamycin, which are expected to occur primarily inthe lipid layers [1]. Clearly, drug penetration is enhancedby occlusion due to the use of petroleum jelly forformulating the drug. Evidently, this also leads to theuptake of rapamycin into the corneocytes, so that analmost continuous drug distribution is observed in thestratum corneum. There is a characteristic drop in drugconcentration near the border to the stratum granulosum,
the top layer of the viable epidermis, marked by greycolor (see Fig. 1 (b) ). This hints that the tight junction
barrier is intact so that the drug cannot penetrateefficiently into the viable epidermis.
Systematic studies involving the primary treatment ofskin with serine protease for simulating the impact ofinflammations have been performed and are currently in
preparation for publication [4].

Fig. 1. Results from STXM of the top skin layers of
fixed human skin: (a) untreated reference skin and (b)skin treated by rapamycin in petroleum jelly. A: opticaldensity at 532 eV; B: assignment of skin regions due
to X-ray absorption characteristics; C: local rapamycin
concentration; D: integrated drug concentration as afunction of depth. The skin surface is marked by anarrow. See text for further details.
[1] K. Yamamoto et al., Anal. Chem. 87 (2015) 6173.
Eur. J. Pharm. Biopharm. 118 (2017) 30.
Eur. J. Pharm. Biopharm. 139 (2019) 68.
[2] G. Germer et al., UVSOR Activity Report 2018 46(2019) 165.
[3] I. N. Koprinarov et al., J. Phys. Chem B 106 (2002)
5358.
[4] G. Germer et al., in preparation (2020).

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Earth and Planetary Sciences

BL4U

Chemical Mapping of Potassium-containing Particles from Residential
Biomass Burning and in Ambient Air
Department of Chemistry and Molecular Biology, University of Gothenburg, 412 96 Gothenburg, Sweden
State Key Joint Laboratory of Environmental Simulation and Pollution Control, College of Environmental
Sciences and Engineering, Peking University, 100871, China
3
UVSOR Synchrotron Facility, Institute for Molecular Science, Okazaki 444-8585, Japan

1
2

Ⅲ- 5

X. Kong1, M. Priestley1, Z. Wu2, T. Ohigashi3, H. Yuzawa3, J. B. C. Pettersson1
and M. Hallquist1

Burning of wood and other biomass fuels for energy
purposes is common on the global scale. Potassium, as
one of the main nutrient elements in plants, is released
during the burning process and it has been considered
as a tracer for biomass burning activities. Here we
employed the STXM technique to visualize potassiumcontaining particles sampled under two circumstances.
One is from a campaign at the Research Institutes of
Sweden (RISE), where a small-scale wood stove was
evaluated regarding its environmental and climate
impacts. The other is from field measurements carriedout in Beijing with a focus on aerosol properties in theurban environment. The STXM results presented in thisstudy were obtained at the BL4U beamline at UVSOR.
At RISE, aerosol samples were collected from theexhaust line of a wood log burning stove. The major
components are soot, organics and inorganic particles.
Figure 1 shows the optical density (OD) of a large
particle aggregate comprised primarily of a large sootstructure with an organic coating that has potassium
particles embedded on its surface. Figure 1e shows thedifference of OD between the photo energy of (c) 295
eV and (d) 297 eV. The dark spots show the locations
where the OD was dramatically enhanced at 297 eV, i.e.
the potassium L3-edge. The potassium salt particles
seem to attach on the boundaries of the aggregate. Soot
is formed in the furnace at high temperatures (>1000 °C), but under these conditions both organic andpotassium compounds are mainly in the gas phase. As
temperature is reduced in the exhaust line, potassium
compounds first nucleate to form potassium salt
particles followed by nucleation and condensation oforganics at lower temperatures. The formed potassiumcontaining particles may agglomerate with existing sootparticles, and they are all coated by organics.
Figure 2 shows an OD image of various types ofaerosol sampled in Beijing, with a photo energy of 300
eV. Several particles were examined in detail with one
example shown here (red square). Full stack scans were
performed on this particle at carbon K-edge (potassium
L-edge), nitrogen K-edge and sulfur L-edge, respectively.
Figures 2 (a)-(c) show the spectra of these three edges onboth the core and coating of the particle. Potassium isfound in the core region but not on the coatings. The
coating mainly contains organic materials. As expectedfrom the chemical complexity of actual atmosphericparticles, all of the ammonium, nitrate [1] and sulphate[2] were found in the core of this single particle,

indicating this is a well-aged and -mixed particle with anorganic coating.

Fig. 1. Optical density of a particle aggregate. The
used photo energies are: (a) 280 eV (pre-edge ofcarbon), (b) 285.5 eV (soot is emphasized), (c) 295 eV(total carbon is emphasized, and also as pre-edge ofpotassium), and (d) 297 eV (potassium is emphasized).
(e) shows the difference between panel (c) and (d).

Fig. 2. OD image of urban particles sampled in Beijing.
An example particle is scanned at (a) carbon K-edge, (b)nitrogen K-edge and (c) sulfur L-edge, respectively. The
spectra measured at the core and on the border weredifferentiated in red and blue respectively.
[1] Török, S., et al., Powder Diffr. 19 (2004) 81.
[2] Jalilehvand, F., Chem. Soc. Rev. 35 (2006) 1256.

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BL4U

Sequestration of Boron in Sediments
Y. Hashimoto1, K. Kobayashi1, H. Suga2 and Y. Takahashi2

Department of Bioapplications and Systems Engineering, Tokyo University of Agriculture and Technology,
Koganei 184-8585, Japan2
Department of Earth and Planetary Science, The University of Tokyo, Tokyo 113-0033, Japan

1

Boron (B) is ubiquitous in the environment, andmajor geogenic sources of B in water include desert,
saline lake, and spring water. Because short-term andlong-term oral exposures to B in animals lead toexpressions of reproductive and developmental
toxicity [1], the elevated levels of B in water supplies
has long been a concern for public health. To lower theenvironmental risk of B contamination in water,
chemical and engineering techniques for removing B
from contaminated water have been developed basedon physicochemical adsorption, coprecipitation, ion
exchange, and reverse osmosis processes. However,
the remaining issues for the current techniques inaqueous B removal reside in the inefficiency in costfor materials and reagents (e.g., resin and flocculation
chemicals) and B removal capacity and kinetics.
Numerous separation materials currently available foraqueous B treatments are less effective for thetreatment of water with high levels of B. Therefore,
there is an urgent need in the development of costeffective methods for the treatment of (waste)waterwith high levels of dissolved B.
The concentrations of B are often high in discharged
water from chemical industries and mining and hot
spring areas. Boron in the aqueous environment wasoften found in minerals including calcite and Fe oxideminerals. Knowledge about how B occurs in theenvironment would help understand the mechanisms of
B sequestration and can be used for developing
technologies for wastewater with high B concentrations.
We examined the distribution of B in sediment collectedfrom a hot spring area with high B conditions using Xray microscopy (STXM)-based
NEXAFS analyses at UVSOR BL4U. A thin sectionof sediment was prepared and analyzed for B on thesurface.
STXM-based merged B images were obtained at theenergy of 188 and 200 eV and their subtraction (Fig.
1). Framboidal pyrite (aspberry-like arrangements ofpyrite grains) was found in the sediment and contained
B with trigonal and tetrahedral coordination. The
distribution of IIIB, IVB and their mixture occupied 4 %,58 %, and 38 %, respectively. This result indicates that
B in this hot spring area occurred mainly with IVB.
Further investigation is needed to reveal the structureof B incorporation to framboidal pyrite.

Ⅲ- 5

Fig. 1. The SEM image about data collecting area for
STXM-based distribution map of boron (a), OD imagesof pre-edge energy (188 eV, b) and post-edge energy(200 eV, c), and boron distribution map subtracted
counts of b from counts of c (d).
[1] WHO (World Health Organization), Guidelines for
Drinking-water Quality: First Addendum to the Fourth
Edition (2017).

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Earth and Planetary Sciences

BL4U

Preliminary Results by a STXM-XANES: Analysis of Small Carry-on
Impactor Debris Operated by the Hayabusa2 Mission
M. Ito1, Y. Takano2, T. Ohigashi3, H. Yuzawa3, Y. Kebukawa4, K. Kiryu4, M. Uesugi5,
N. Tomioka1, Y. Kodama6, H. Naraoka7, T. Yada8 and M. Abe8
Kochi Institute for Core Sample Research, JAMSTEC, Nankoku 783-8502, Japan,
2
Biogeochemistry Program, JAMSTEC, Yokosuka 237-0061, Japan,
3
UVSOR Synchrotron Facility, Institute for Molecular Science, Okazaki 444-8585, Japan,
4
Yokohama National University, Yokohama 240-8501, Japan,
5
JASRI/SPring-8, Sayo-cho 679-5198, Japan,
6
Marine Works Japan, Yokosuka 237-0063, Japan,
7
Kyushu University, Fukuoka 819-0395, Japan,
8
JAXA, Sagamihara 252-5210, Japan1

Ⅲ- 5

The JAXA Hayabusa2 mission is a combination ofan asteroidal sample return and detailed spectroscopic
observations of the C-type asteroid Ryugu for
understanding of the Solar System evolution from thepoint of view of organics, and water as hydrous
minerals [1]. The Hayabusa2 spacecraft was thought
to be successfully obtained the Ryugu samples (anexpected total amount of ~ 100 mg) from surface andsubsurface on Feb. and Apr. 2019 and will return to the
Earth on Dec. 2020.
To obtain the subsurface samples of the Ryugu, Small
Carry-on Impactor (SCI) [2-4] was used to generate an
artificial crater on the surface. A force of the SCI was
made
by
HMX
(High-Melting
Explosive;
cyclotetramethylenetetranitramine; C4H8N8O8, 296.15)
together with HTPB (Hydroxyl TerminatedPolybutadiene), IDP (Isodecyl palargonate; C19H38O2)
and IPDI (Isophorone diisocyanate; C12H18N2O2) [3,4].
Takano and coworkers [5] reported analytical results of
SEM-EDS, TD-GC/MS, SPME-GC/MS, and isotope
mass spectrometry for N and C isotopes. They foundvolatiles of aliphatic and aromatic carbon structures,and the several functional groups (hydroxyl, aldehyde,
nitrile, and carboxyl) [5].
The explosion products generated by their chemical
reactions with HMX, HTPB, IDP and IPDI should be
investigated prior to analysis of the Ryugu samples
because those materials might be possible contaminantsin the subsurface samples in the sample container. In
fact, Matsuoka and coworkers pointed out that the low
albedo of the asteroid Ryugu could be explained by a
combination of C-rich material, grains size, porosity
and space weathering effects on the asteroid surface
materials [6]. A study of the optical navigation camera
on Hayabusa2 space craft reported that the asteroid
Ryugu was composed of fluffy cosmic dust particles
which is like the carbonaceous materials [7].
It is, therefore, necessary to analyze the SCI
fragments and its products for comparison to the
Ryugu samples. These results will provide relevant
information to the initial analysis in advance thearrival of the Ryugu samples to the Earth.
In this study, we have conducted STXM-XANES

analysis of the experimental explosion-products of the
HMX mixture (Fig. 1) for sub-micrometer-scale
functional groups of carbon, nitrogen and oxygen. Eachfragment shows similar characteristics in O, N but some
exceptions of C-XANES in sub-micrometer scale.
To acquire additional chemical properties of the
products, we have carried out series of analysis
utilizing an optical microscope for micrometer-scale
texture, SEM-EDS for texture, microFT-IR for
functional groups, micro-Raman for structures ofcarbonaceous materials. In the future, we will measurehydrogen, carbon and nitrogen isotopes using the
JAMSTEC NanoSIMS (nano-scale Secondary Ion
Mass Spectroscopy) for further characterizations of
the samples.

Fig. 1. Experimental products of HMX mixture.

[1] Tachibana et al., Geochemical Journal 48 (2014) 571.
[2] Arakawa et al., Science 368 (2020) 67.
[3] Saiki et al., Acta Astronautica 84 (2013) 227.
[4] Saiki et al., Space Science Reviews 208 (2016) 165.
[5] Takano et al., (2014) in Hayabusa 2014: 2nd Symp.
Solar System Materials, P03.
[6] Matsuoka et al. (2019) 50th LPSC (LPI Contrib.
No. 2132), abstract#1534.
[7] Okada et al., Nature 579 (2020) 518.

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Characterization of Lysosomal Storage Diseases Using STXM
T. Mansikkala1,2, T. Ohigashi3, H. Yuzawa3, M. Patanen1,2, I. Miinalainen2, S. M. Kangas4,
A. E. Hiltunen4, E.-V. Immonen1,4, N. Kosugi3, R. Hinttala4, J. Uusimaa4 and M. Huttula1
Nano 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
PEDEGO Research Unit and Medical Research Center Oulu, and Oulu University Hospital, PO Box 5000,
90014 University of Oulu, Finland
1

In this work, we have used scanning transmission Xray microscopy (STXM) technique at BL4U beamline
to study patient samples with lysosomal storage
diseases and cultured patient-derived fibroblasts with amutation in NHLRC2 gene. The NHLRC2 encodes an
NHL repeat containing protein 2 ubiquitously present invarious types of tissues. Mutations in NHLRC2 gene
have been linked to fatal cerebropulmonary disease inchildren with fibrosis, neurodegeneration and cerebral
angiomatosis (FINCA disease), but the functional roleof the NHLRC2 is still unknown [1]. Transmission
Electron Microscopy (TEM) analysis of immortalized
cell cultures from FINCA patients demonstrated
multilamellar and multivesicular bodies of unknown
cellular origin, and it is suggested that NHLRC2
dysfunction enhances tissue fibrosis [2]. Previously, we
reported an STXM study of FINCA patient tissue
samples which were in line with TEM results [3].
Our aim was to investigate samples from known
lysosomal storage diseases and FINCA disease. Muscle
and kidney tissues samples were used, and they had
clear enlarged lysosome clusters typical for lysosomal
storage diseases. We also had cultured FINCA patientderived fibroblasts to test if cell culture samples can beimaged. The tissue samples were fixed in 4 %paraformaldehyde and 2.5 % glutaraldehyde in 0.1 M
phosphate-buffered saline. Prior to embedding in resinsome samples were stained with 1 % OsO4 and/or
uranyl acetate. The 100 nm and 150 nm thick sectionsof resin-embedded sample were imaged at Biocenter
Oulu using a Tecnai G2 Spirit 120 kV TEM (FEI,
Eindhoven, The Netherlands) equipped with a Quemesa
CCD camera (Olympus Soft Imaging Solutions GmbH,
Münster, Germany) to find lysosome clusters on thesamples. The same area was then measured with STXM
from the subsequent thin sections.
Figure 1 shows example of lysosomes in muscle
tissue from patient with a lysosomal storage disease. A
cluster analysis of STXM image stacks (Fig. 1 (b)) was
performed using a MANTIS software [4]. The cluster
analysis was able to differentiate cell components rather
consistently based on their spectral information. The
spectra obtained at C K-edge are very similar in eachcluster as can be seen in Fig. 1 (c). This is caused by theresin that penetrates throughout the tissue andcontributes mostly to the spectra. There seems to be nodifference between the lysosomes and empty areas
filled with resin (blue color in Fig. 1 (b)), indicating thatthe material inside lysosomes is lost during the
preparation process.
Even though the resin fills the sample, its effect on
the spectra can be reduced by removing the resin
spectrum from the data. Figure 2 shows an example
where the resin spectrum is subtracted with a factor of
0.9 using aXis2000. The exact multiplier is dependent
on how much the embedding material penetrates the cellstructures and it can vary. By removing the resin, we candetect spectral features that would otherwise be masked
by the resin. In Fig. 2 we can see that there is strong
peak at 288 eV differing from the typical peak for resin
at 288.4 eV. This means that even though it is not ideal,
we can study resin embedded samples and provide
valuable data from them. However, the goal ofidentifying the storage material within lysosomes was
not achieved.

Fig. 1. (a) TEM image of the lysosome cluster frommuscle tissue of a patient with lysosomal storage
disease. (b) Cluster analysis of lysosomes imaged with
STXM from same area as (a) but from a different thinslice. (c) Spectra of the clusters in (b).

Fig. 2. Locations of fitted spectra in muscle tissue, (a)collagen, (b) lysosomes and (c) muscle fibers. (d)Spectra of the structures, red is collagen (a), green islysosomes (b) and blue is muscle fibers (c). Collagenfiber bundle is marked with *, lysosomes are marked
with Δ and muscle fiber with ↑. Scale bar is 1 μm.
[1] J. Uusimaa et al., Acta Neuropathol. 135 (2018) 727.
[2] T. Paakkola et al., Hum. Mol. Gen. 27 (2018) 4288.
[3] E.-V. Immonen et al., UVSOR Activity Report2017 45 (2018) 161.
[4] M. Lerotic et al., J. Synchr. Rad. 21 (2014) 1206.

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UVSOR User 1