Protocol

In Situ Desorption Electrospray Ionization (DESI) Analysis of Tissue Sections

This protocol was adapted from "Desorption Electrospray Ionization: Proteomics Studies by a Method that Bridges ESI and MALDI," Chapter 6, in Proteomics: Methods Express (eds. O’Connor and Hames). Scion Publishing Ltd., Oxfordshire, UK, 2007.

INTRODUCTION

Desorption electrospray ionization (DESI) allows in situ analysis of biological tissues. The analysis of less abundant protein constituents within a tissue sample often requires the removal of lipid species prior to analysis, similar to the situation with matrix-assisted laser desorption/ionization (MALDI). After removal of lipid constituents, the tissue can be treated with protease to degrade proteins present in the tissue. The tryptic products can be investigated directly from the tissue using DESI. The spectra obtained feature ions of tryptic fragments from abundant proteins present in the tissue sample. The digestion is usually not complete; hence, the presence of missed cleavage sites is typical in the peptides detected. The signal is more stable for longer times than in the case of deposited samples, so the recording of mass spectrometry (MS)/MS data is simple in this case. DESI-MS is an emerging technique with great promise, but its application range is still being investigated. Therefore, the protocol for DESI-MS analysis of tissue sections presented here provides general procedures used for the applications that have been investigated so far. Optimal ion source parameters and surface types may vary depending on the application.