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Device-Controlled Microcondensation for Spatially Confined On-Tissue Digests in MALDI Imaging of N -Glycans

Affiliation
Center for Mass Spectrometry and Optical Spectroscopy (CeMOS), Mannheim University of Applied Sciences, Paul-Wittsack-Str. 10, 68163 Mannheim, Germany
Fülöp, Annabelle;
ORCID
0000-0002-7564-1385
Affiliation
Center for Mass Spectrometry and Optical Spectroscopy (CeMOS), Mannheim University of Applied Sciences, Paul-Wittsack-Str. 10, 68163 Mannheim, Germany
Marsching, Christian;
Affiliation
SunChrom Wissenschaftliche Geräte GmbH, 61381 Friedrichsdorf, Germany
Barka, Frederik;
Affiliation
Center for Mass Spectrometry and Optical Spectroscopy (CeMOS), Mannheim University of Applied Sciences, Paul-Wittsack-Str. 10, 68163 Mannheim, Germany
Ucal, Yasemin;
ORCID
0000-0002-0945-678X
Affiliation
Metabolic Crosstalk in Cancer, German Consortium of Translational Cancer Research (DKTK), German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany
Pfänder, Pauline;
ORCID
0000-0001-5575-9821
Affiliation
Metabolic Crosstalk in Cancer, German Consortium of Translational Cancer Research (DKTK), German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany
Opitz, Christiane A.;
Affiliation
SunChrom Wissenschaftliche Geräte GmbH, 61381 Friedrichsdorf, Germany
Barka, Günes;
ORCID
0000-0003-0802-6451
Affiliation
Center for Mass Spectrometry and Optical Spectroscopy (CeMOS), Mannheim University of Applied Sciences, Paul-Wittsack-Str. 10, 68163 Mannheim, Germany
Hopf, Carsten

On-tissue enzymatic digestion is a prerequisite for MALDI mass spectrometry imaging (MSI) and spatialomic analysis of tissue proteins and their N -glycan conjugates. Despite the more widely accepted importance of N -glycans as diagnostic and prognostic biomarkers of many diseases and their potential as pharmacodynamic markers, the crucial sample preparation step, namely on-tissue digestion with enzymes like PNGaseF, is currently mainly carried out by specialized laboratories using home-built incubation arrangements, e.g., petri dishes placed in an incubator. Standardized spatially confined enzyme digests, however, require precise control and possible regulation of humidity and temperature, as high humidity increases the risk of analyte dislocation and low humidity compromises enzyme function. Here, a digestion device that controls humidity by cyclic ventilation and heating of the slide holder and the chamber lid was designed to enable controlled micro-condensation on the slide and to stabilize and monitor the digestion process. The device presented here may help with standardization in MSI. Using sagittal mouse brain sections and xenografted human U87 glioblastoma cells in CD1 nu/nu mouse brain, a device-controlled workflow for MALDI MSI of N -glycans was developed.

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