Tissue protein imaging at 1 μm laser spot diameter for high spatial resolution and high imaging speed using transmission geometry MALDI TOF MS
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We have achieved protein imaging mass spectrometry capabilities at sub-cellular spatial resolution and at high acquisition speed by integrating a transmission geometry ion source with time of flight mass spectrometry. The transmission geometry principle allowed us to achieve a 1-μm laser spot diameter on target. A minimal raster step size of the instrument was 2.5 μm. Use of 2,5-dihydroxyacetophenone robotically sprayed on top of a tissue sample as a matrix together with additional sample preparation steps resulted in single pixel mass spectra from mouse cerebellum tissue sections having more than 20 peaks in a range 3–22 kDa. Mass spectrometry images were acquired in a standard step raster microprobe mode at 5 pixels/s and in a continuous raster mode at 40 pixels/s.
KeywordsMALDI Transmission geometry High spatial resolution MS imaging Protein imaging
The authors would like to thank George Mills (SimulTOF Systems) and Boone Prentice and other members of the Vanderbilt Mass Spectrometry Research Center. This project was supported by grants from National Institutes of Health National Institute of General Medical Sciences NIH/NIGMS P41 GM103391-04 and NIH/NIGMS R01 GM058008-15.
- 10.SimulTOF 200 Combo | SimulTOF Systems. http://www.simultof.com/content/simultof-200-combo.
- 11.Ritzau S, Hayden K, Vestal M (2012) Improved MALDI-TOF performance with practical implementation of very high post-acceleration. In: 60th ASMS Conference on Mass Spectrometry and Allied Topics, Vancouver, CanadaGoogle Scholar
- 12.Trim P, Djidja M-C, Atkinson S, Oakes K, Cole L, Anderson DG, Hart P, Francese S, Clench M (2010) Introduction of a 20 kHz Nd:YVO4 laser into a hybrid quadrupole time-of-flight mass spectrometer for MALDI-MS imaging. Anal Bioanal Chem 397(8):3409–3419. doi: 10.1007/s00216-010-3874-6 CrossRefGoogle Scholar
- 15.Yang J, Zavalin A, Caprioli R (2014) Highly robust sample preparation with 2,5-dihydroxyacetophenone for MALDI imaging of proteins (2 -70 kDa) at high spatial resolution (5 μm). In: 62th ASMS Conference on Mass Spectrometry and Allied Topics, Baltimore, MDGoogle Scholar
- 16.Wenzel T, Sparbier K, Mieruch T, Kostrzewa M (2006) 2,5-Dihydroxyacetophenone: a matrix for highly sensitive matrix-assisted laser desorption/ionization time-of-flight mass spectrometric analysis of proteins using manual and automated preparation techniques. Rapid Commun Mass Spectrom 20(5):785-789. doi: 10.1002/rcm.2378
- 18.Prophet EB, Armed Forces Institute of Pathology et al (1992) Laboratory methods in histotechnology. American Registry of Pathology, WashingtonGoogle Scholar