Laser Ablation with Vacuum Capture for MALDI Mass Spectrometry of Tissue

© 2015 American Society for Mass Spectrometry. We have developed a laser ablation sampling technique for matrix-assisted laser desorption ionization (MALDI) mass spectrometry and tandem mass spectrometry (MS/MS) analyses of in-situ digested tissue proteins. Infrared laser ablation was used to remove biomolecules from tissue sections for collection by vacuum capture and analysis by MALDI. Ablation and transfer of compounds from tissue removes biomolecules from the tissue and allows further analysis of the collected material to facilitate their identification. Laser ablated material was captured in a vacuum aspirated pipette-tip packed with C18 stationary phase and the captured material was dissolved, eluted, and analyzed by MALDI. Rat brain and lung tissue sections 10 μm thick were processed by in-situ trypsin digestion after lipid and salt removal. The tryptic peptides were ablated with a focused mid-infrared laser, vacuum captured, and eluted with an acetonitrile/water mixture. Eluted components were deposited on a MALDI target and mixed with matrix for mass spectrometry analysis. Initial experiments were conducted with peptide and protein standards for evaluation of transfer efficiency: a transfer efficiency of 16% was obtained using seven different standards. Laser ablation vacuum capture was applied to freshly digested tissue sections and compared with sections processed with conventional MALDI imaging. A greater signal intensity and lower background was observed in comparison with the conventional MALDI analysis. Tandem time-of-flight MALDI mass spectrometry was used for compound identification in the tissue

Particle Formation in Ambient MALDI Plumes

The ablated particle count and size distribution of four solid matrix materials commonly used for matrix-assisted laser desorption ionization (MALDI) were measured with a scanning mobility particle sizer (SMPS) combined with a light scattering aerodynamic particle sizer (APS). The two particle sizing instruments allowed size measurements in the range from 10 nm to 20 μm. The four solid matrixes investigated were 2,5-dihydroxybenzoic acid (DHB), 4-nitroaniline (NA), α-cyano-4-hydroxycinnamic acid (CHCA), and sinapic acid (SA). A thin film of the matrix was deposited on a stainless steel target using the dried droplet method and was irradiated with a 337 nm nitrogen laser at atmospheric pressure. The target was rotated during the measurement. A large number of nanoparticles were produced, and average particle diameters ranged from 40 to 170 nm depending on the matrix and the laser fluence. These particles are attributed to agglomeration of smaller particles and clusters and/or hydrodynamic sputtering of melted matrix. A coarse particle component of the distribution was observed with diameters between 500 nm and 2 μm. The coarse particles were significantly lower in number but had a total mass that was comparable to that of the nanoparticles. The coarse particles are attributed to matrix melting and spallation. Two of the compounds, CHCA and SA, had a third particle size distribution component in the range of 10 to 30 nm, which is attributed to the direct ejection of clusters. © 2011 American Chemical Society