Enhancement of protein sensitivity for MALDI imaging mass spectrometry after chemical treatment of tissue sections.
MALDI imaging mass spectrometry (IMS) has become a valuable tool for the investigation of the content and distribution of molecular species in tissue specimens. Numerous methodological improvements have been made to optimize tissue section preparation and matrix deposition protocols, as well as MS data acquisition and processing. In particular for proteomic analyses, washing the tissue sections before matrix deposition has proven useful to improve spectral qualities by increasing ion yields and the number of signals observed. We systematically explore here the effects of several solvent combinations for washing tissue sections. To minimize experimental variability, all of the measurements were performed on serial sections cut from a single mouse liver tissue block. Several other key steps of the process such as matrix deposition and MS data acquisition and processing have also been automated or standardized. To assess efficacy, after each washing procedure the total ion current and number of peaks were counted from the resulting protein profiles. These results were correlated to on-tissue measurements obtained for lipids. Using similar approaches, several selected washing procedures were also tested for their ability to extend the lifetime as well as revive previously cut tissue sections. The effects of these washes on automated matrix deposition and crystallization behavior as well as their ability to preserve tissue histology were also studied. Finally, in a full-scale IMS study, these washing procedures were tested on a human renal cell carcinoma biopsy.
Seeley
Erin H
EH
Mass Spectrometry Research Center and Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, TN 37232, USA.
Oppenheimer
Stacey R
SR
Mi
Deming
D
Chaurand
Pierre
P
Caprioli
Richard M
RM
D000818:Animals; D002292:Carcinoma, Renal Cell / Q000473:pathology; D006801:Humans; D007202:Indicators and Reagents; D007680:Kidney Neoplasms / Q000473:pathology; D008099:Liver / Q000473:pathology; D051379:Mice; D011506:Proteins / Q000737:chemistry*; D012997:Solvents; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D007202:Indicators and Reagents; D011506:Proteins; D012997:Solvents