XIV
h
International Conference on Molecular Spectroscopy, Białka Tatrzańska 2017
70
T2: O–2
2D Raman study of the healthy and epileptic rat brain tissue
Julia Sacharz
1
, Aleksandra Wesełucha-Birczyńska
1
, Janina Zięba-Palus
2
,
Marian. H. Lewandowski
3
, Rafał Kowalski
2
, Katarzyna Palus
3
, Łukasz Chrobok
3
1
Faculty of Chemistry, Jagiellonian University, Krakow, Poland,
e-mail: julia.sacharz@doctoral.uj.edu.pl
2
Institute of Forensic Research, Krakow, Poland
3
Department of Neurophysiology and Chronobiology, Institute of Zoology, Jagiellonian University,
Krakow, Poland
The aim our work is to recognize if Raman spectroscopy can detect brain abnormalities
responsible for epilepsy at the molecular level. Absence epilepsy is the neurological disorder
characterized by the pathological spike-and wave discharges present in the
electroencephalogram, accompanying a sudden loss of consciousness [1]. Experiments were
performed on male rats from WAG/Rij strain, as it is the well-established model of epilepsy [2]
and on control group of Wistar rats. Three differrent brain areas of the rats brain tissue were
studied: the somatosensory cortex (Sc), the dorsal lateral geniculate nucleus of the thalamus
(DLG) and the cerebellar cortex (Cc). The Raman spectra of the fresh brain scraps kept in
artificial cerebrospinal fluid were collected using as an excitation source 442 nm, 785 nm, 514.5
nm and 1064 nm laser lines. The rat brain is composed of over a thousand functionally and
anatomically independent structures [3, 4], which makes it challenge to study. The differences in
electrophysiological functioning of Sc and DLG brain structures – the presumable origin of
absence seizures, in WAG/Rij rats were indicated [2]. 2D correlation analysis, performed using
the Noda method, allowed to identify the subtle differences in tissue structure connected with
brain pathology [5]. Average Raman spectra were used as input data, while the laser wavelength
was regarded as an external perturbation in 2D correlation (Fig. 1).
Fig. 1. 2D asynchronous maps for Sc, DLG and Cc brain areas in the 1735–1600 cm
–1
range showing differences
in the spectra between healthy and epileptic brain structures.
Keywords: Epilepsy, Raman Spectroscopy, 2D Correlation
References
[1] V. Crunelli, N. Leresche, Nature Reviews Neuroscience 3 (2002) 371.
[2] T. Budde, J.R. Huguenard, Models of Seizures and Epilepsy (2006) 73.
[3] T.S. Reddy, R. Rajalakshmi, C.V. Ramakrishnan, Int. J. Developmental Neuroscience 1 (1983) 65.
[4] A. Wesełucha-Birczyńska, J. Sacharz, J. Zięba-Palus, M.H. Lewandowski, R. Kowalski, K. Palus, L.
Chrobok, M. Birczyńska, A. Sozańska, Vib. Spectrosc 85 (2016) 48.
[5] I. Noda, Appl. Spectrosc. 47 (1993) 1329.
W
is
ta
Sc
DL
Cc
W
A
G
/R
ij
XIV
h
International Conference on Molecular Spectroscopy, Białka Tatrzańska 2017
71
T2: O–3
Is SR-FTIR spectroscopy has a potential
as a tissue classifier in ovarian cancer?
Maria Magdalena Grzelak
1
, Marek Lankosz
1
, Z. Stęgowski
1
, Sylwia Hotloś
1
,
Łukasz Chmura
2
, and Dariusz Adamek
3
1
AGH University of Science and Technology, Faculty of Physics and Applied Computer Science,
al. A. Mickiewicza 30, 30-059, Krakow,Poland, e-mail: Maria.Grzelak@fis.agh.edu.pl
2
Chair of Pathomorphology, Faculty of Medicine, Jagiellonian University, ul. Grzegorzecka 16,
31-531 Krakow, Poland
Ovarian cancer is the seventh most common cancer in women worldwide (18 most common
cancer overall). Neoplasms tissues derived from epithelial ovarian surface are heterogeneous
morphology group, including wide spectrum of clinical signs and the wide histopathological
scope. Currently available methods and test do not provide the required sensitivity and
specificity for accurate diagnosis of the ovarian cancer. There is need to find detection tool to
diagnosis this type of cancer. Infrared (IR) spectroscopy of the tissue is a rapid, versatile, and
relatively non-invasive approach which could characterize biomolecular alterations due to
cancer and has potential to be utilized as a screening or the diagnostic tool [1].
The aim of this study was to check if some biomolecules can be treated as ovarian cancer
bio-indicator and to optimize the procedure of samples preparation for the measurement:
thickness and deposit material. Another aspect of this study was to check if any differences or
similarities in the distribution of the molecules can be find in neoplastic and control area of the
tissue.
The samples designed to chemical and molecular analysis were taken intraoperatively from
ovarian tumors of different type and degrees of malignancy. The samples were collected from
12 patients in different age. The samples after the surgery were frozen in -80oC. Tissue material
for the experiment was cryo-sectioned to slices of 5 μm thick, mounted on a square silicon
nitride membrane window (2x2 mm, 200 nm thick) on silicon frame or of 20 μm thick, mounted
on barium fluoride disc (13 mm diameter, 2 mm thick) and dried in vacuum. The experiment
were performed at the beamline ID2 (ESRF, Grenoble, France) The absorption spectra were
collected in trans-reflective mode using the infrared microscope (Continuum Thermo Nicolet)
coupled with the Thermo Nicolet Nexus spectrometer.
Set of biomolecules (amid massive, lipid massive and fingerprint region) in neoplastic and
control area of the tissues were examined. Distributions of biomolecules were presented on 2D
maps. The molecular structure mainly for determining the secondary structure of proteins were
also examined in regions of the amide massive. [2]. The differences in the IR spectra and
distribution of molecules for various types of ovarian tumors were observed. Statistical analysis
of the measurement data was carried out with the STATISTICA package. The Mann Whitney U
test and cluster analysis were applied for statistical evaluation.
Keywords: ovarian cancer; chemical imaging; SR-FTIR; statistical analysis
Acknowledgment
We acknowledge the European Synchrotron Radiation Facility for provision of synchrotron radiation
facilities and we would like to thank Dr M. Salome, Dr H. Castillo-Michel, Dr B. Hesse and Dr G. Veronesi
for assistance in using beamline ID21. The research leading to these results has received funding from
ESRF and the Ministry of Science and Higher Education (Warsaw, Poland)
References
[1] K. Gajjar, J. Trevisan, G. Owens, P.J. Keating, N.J. Wood, H.F. Stringfellow, P.L. Martin-Hirsch, F.
Martin, Analyst 138(14) (2013) 3917.
[2] C. Petibois, G. Deleris , Trends in Biotehnology 24(10) (2006) 455.
Dostları ilə paylaş: |