Essential tactics of tissue preparation and matrix nano-spotting for successful compound imaging mass spectrometry.
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 (1alpha,2beta,4beta,7beta)-7-[(hydroxidi-2-thienylacetyl)oxy]-9,9-dimethyl-3-oxa-9-azoniatricyclo[3.3.1.0(2,4)]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.
Végvári
Akos
A
Clinical Protein Science & Imaging, Dept. of Measurement Technology and Industrial Electrical Engineering, Lund University, BMC C13, SE-221 84 Lund, Sweden.
Fehniger
Thomas E
TE
Gustavsson
Lena
L
Nilsson
Anna
A
Andrén
Per E
PE
Kenne
Kerstin
K
Nilsson
Johan
J
Laurell
Thomas
T
Marko-Varga
György
G
D000818:Animals; D001993:Bronchodilator Agents / Q000494:pharmacology; D003460:Crystallization; D001239:Inhalation; D008168:Lung / Q000473:pathology; D008297:Male; D008856:Microscopy, Fluorescence / Q000379:methods; D036103:Nanotechnology / Q000379:methods*; D020543:Proteome; D040901:Proteomics / Q000295:instrumentation* / Q000379:methods*; D051381:Rats; D017208:Rats, Wistar; D012602:Scopolamine Derivatives / Q000494:pharmacology; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000295:instrumentation / Q000379:methods*; D000069447:Tiotropium Bromide
D001993:Bronchodilator Agents; D020543:Proteome; D012602:Scopolamine Derivatives; D000069447:Tiotropium Bromide