Quantitative imaging mass spectrometry of renal sulfatides:
validation by classical mass spectrometric methods |
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Authors: | Christian Marsching Richard Jennemann Raphael Heilig Hermann-Josef Gr?ne Carsten Hopf Roger Sandhoff |
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Institution: | 2. Lipid Pathobiochemistry Group within German Cancer Research Center (DKFZ), Heidelberg, Germany;4. Department of Cellular and Molecular Pathology, German Cancer Research Center (DKFZ), Heidelberg, Germany;11. Instrumental Analytics and Bioanalytics, Mannheim University of Applied Sciences, Mannheim, Germany |
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Abstract: | Owing to its capability of discriminating subtle mass-altering structural
differences such as double bonds or elongated acyl chains, MALDI-based imaging
MS (IMS) has emerged as a powerful technique for analysis of lipid distribution
in tissue at moderate spatial resolution of about 50 μm. However, it is
still unknown if MS1-signals and ion intensity images correlate with
the corresponding apparent lipid concentrations. Analyzing renal sulfated
glycosphingolipids, sulfatides, we validate for the first time IMS-signal
identities using corresponding sulfatide-deficient kidneys. To evaluate the
extent of signal quenching effects interfering with lipid quantification, we
surgically dissected the three major renal regions (papillae, medulla, and
cortex) and systematically compared MALDI IMS of renal sulfatides with
quantitative analyses of corresponding lipid extracts by on-target MALDI
TOF-MS and by ultra-performance
LC-ESI-(triple-quadrupole)tandem MS. Our results demonstrate a generally strong
correlation (R2 > 0.9) between the local relative sulfatide
signal intensity in MALDI IMS and absolute sulfatide quantities determined by
the other two methods. However, high concentrations of sulfatides in the
papillae and medulla result in an up to 4-fold signal suppression. In
conclusion, our study suggests that MALDI IMS is useful for semi-quantitative
dissection of relative local changes of sulfatides and possibly other lipids in
tissue. |
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Keywords: | liquid chromatography matrix-assisted laser desorption/ionization-time-of-flight electrospray ionization-mass spectrometry tandem mass spectrometry galactosylceramide I3-sulfate lactosylceramide II3-sulfate cortex medulla papillae |
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