Essential tactics of tissue preparation and matrix nano-spotting for successful compound imaging mass spectrometry |
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Authors: | Ákos Végvári Thomas E. Fehniger Lena Gustavsson Anna Nilsson Per E. Andrén Kerstin Kenne Johan Nilsson Thomas Laurell György Marko-Varga |
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Affiliation: | 1. Clinical Protein Science & Imaging, Dept. of Measurement Technology and Industrial Electrical Engineering, Lund University, BMC C13, SE-221 84 Lund, Sweden;2. AstraZeneca R&D Lund, Scheelevägen 8, SE-221 87 Lund, Sweden;3. Dept. of Pharmaceutical Biosciences, Medical Mass Spectrometry, Uppsala University, BMC, P.O. Box 583, SE-751 23 Uppsala, Sweden;4. AstraZeneca R&D Södertälje, Safety Assessment, SE-151 85 Södertälje, Sweden |
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Abstract: | The ultimate goal of MALDI-Imaging Mass Spectrometry (MALDI-IMS) is to achieve spatial localization of analytes in tissue sections down to individual tissue compartments or even at the level of a few cells. With compound tissue imaging, it is possible to track the transportation of an unlabelled, inhaled reference compound within lung tissue, through the application of MALDI-IMS. The procedure for isolation and preparation of lung tissues is found to be crucial in order to preserve the anatomy and structure of the pulmonary compartments.To avoid delocalization of analytes within lung tissue compartments we have applied an in-house designed nano-spotter, based on a microdispenser mounted on an XY table, of which movement and spotting functionality were fully computer controlled. We demonstrate the usefulness of this platform in lung tissue sections isolated from rodent in vivo model, applied to compound tissue imaging as exemplified with the determination of the spatial distribution of (1α,2β,4β,7β)-7-[(hydroxidi-2-thienylacetyl)oxy]-9,9-dimethyl-3-oxa-9-azoniatricyclo[3.3.1.02,4]nonane, also known as tiotropium. We provide details on tissue preparation protocols and sample spotting technology for successful identification of drug in mouse lung tissue by using MALDI-Orbitrap instrumentation. |
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