Balkan Journal of Medical Genetics

RENOPATHOLOGICAL MICROSTRUCTURE VISUALIZATION FROM FORMALIN FIXED KIDNEY TISSUE BY MATRIXASSISTED LASER/DESORPTION IONIZATION-TIME-OFFLIGHT MASS SPECTROMETRY IMAGING
Fröhlich S1, Putz B1, Schachner H2, Kerjaschki D2,Allmaier G1, Marchetti-Deschmann M1,*
*Corresponding Author: Dr. Martina Marchetti-Deschmann, Vienna University of Technology, Institute of Chemical Technologies and Analytics, Getreidemarkt 9/164-IAC, 1060 Vienna, Austria; Tel.: +43-1- 58801-15152; Mobile: +43+664-605887663; Fax: +43-1-58801-915162; E-mail: martina.marchettideschmann@ tuwien.ac.at
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INTRODUCTION

A combination of histopathology and mass spectrometry imaging (MSI) offers comprehensive information regarding structure, molecular composition and pathological information in tissue samples. Mass spectrometry imaging is a rapidly developing technique using spatially resolved mass spectrometry (MS) techniques to simultaneously trace distributions of hundreds of biomolecules directly from tissue samples using essentially the same technology. Peptides, proteins, pharmaceuticals and metabolites can also be analyzed but without a label and without prior knowledge. With MSI, molecular peak information is correlated to the underlying tissue architecture and a virtual image is rebuilt with respect to the intensity of each molecular species to understand the distribution of differential signals. Renal dysfunction has a high demand for early stage identification to initiate healing processes. Recent investigations proved nephropathy is associated with insulin resistance leading to malfunctions of the glomerular filtration [1], manifested in potential glomerular basement membrane modifications [2]. Lipid phosphatase levels, promoting podocyte apoptosis leading to diabetic nephropathy, are upregulated before histological changes are observed [1]; moreover, the membrane contains protein complexes associated with cholesterol binding which alters the lipid environment [2]. Lipid accumulations have been reported for diabetic kidney diseases [3] and oxidized phosphatidylcholine species seem to be associated with renal dysfunction [4]. Recently tubuli-related phosphatidylcholine classes identified by MSI were correlated to immunoglobulin A nephropathy [5]. Thus, for visualizing renopathological microstructures, MSI is a promising tool, allowing further discoveries of yet unknown disease-related analytes. Special challenges regarding ion suppression effects during the matrix-assisted laser desorption/ ionization (MALDI) process were observed whilst obtaining lipidomic information using MSI. Consequently, special attention had to be paid to sample preparation methods, regarding washing procedures, matrix application and protein denaturation, making adaptation to the particular analytical question mandatory [6].

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