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  Sampling of Tissues with Laser Ablation for Proteomics: Comparison of Picosecond Infrared Laser and Microsecond Infrared Laser

Krutilin, A., Maier, S., Schuster, R., Kruber, S., Kwiatkowski, M., Robertson, W., et al. (2019). Sampling of Tissues with Laser Ablation for Proteomics: Comparison of Picosecond Infrared Laser and Microsecond Infrared Laser. Journal of Proteome Research, 18(3), 1451-1457. doi:10.1021/acs.jproteome.9b00009.

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Item Permalink: http://hdl.handle.net/21.11116/0000-0001-DDC0-D Version Permalink: http://hdl.handle.net/21.11116/0000-0004-94EE-9
Genre: Journal Article

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 Creators:
Krutilin, A.1, 2, Author              
Maier, S.1, 2, Author              
Schuster, R.3, Author
Kruber, S.1, 2, Author              
Kwiatkowski, M.4, Author
Robertson, W.1, 2, Author              
Miller, R. J. D.1, 2, 5, Author              
Schlüter, H.6, Author
Affiliations:
1Miller Group, Atomically Resolved Dynamics Department, Max Planck Institute for the Structure and Dynamics of Matter, Max Planck Society, ou_1938288              
2Center for Free Electron Laser Science, ou_persistent22              
3University of Hamburg, ou_persistent22              
4Groningen Research Institute of Pharmacy, Pharmacokinetics, Toxicology and Targeting, University of Groningen, ou_persistent22              
5Departments of Chemistry and Physics, University of Toronto, ou_persistent22              
6Institute of Clinical Chemistry and Laboratory Medicine, University Medical Center Hamburg-Eppendorf, ou_persistent22              

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Free keywords: laser ablation; mass spectrometry; proteomics; tissue sampling and homogenization
 Abstract: It was recently shown that sampling of tissues with a picosecond infrared laser (PIRL) for analysis with bottom-up proteomics is advantageous compared to mechanical homogenization. Because the cold ablation of tissues with PIRL irradiation is soft, proteins remain intact and even enzymatic activities are detectable in PIRL homogenates. In contrast, it was observed that irradiation of tissues with a microsecond infrared laser (MIRL) heats the tissue, thereby causing significant damage. In this study, we investigated the question if sampling of tissues with a MIRL for analysis of their proteomes via bottom-up proteomics is possible and how the results are different from sampling of tissues with a PIRL. Comparison of the proteomes of the MIRL and PIRL tissue homogenates showed that the yield of proteins identified by bottom-up proteomics was larger in PIRL homogenates of liver tissue, whereas the yield was higher in MIRL homogenates of muscle tissue, which has a significantly higher content of connective tissue than liver tissue. In the PIRL homogenate of renal tissue, enzymatic activities were detectable, whereas in the corresponding MIRL homogenate, enzymatic activities were absent. In conclusion, MIRL and PIRL pulses are suited for sampling tissues for bottom-up proteomics. If it is important for bottom-up proteomic investigations to inactivate enzymatic activities already in the tissue before its ablation, MIRL tissue sampling is an option, because the proteins in the tissues are denatured and inactivated by the heating of the tissue during irradiation with MIRL irradiation prior to the ablation of the tissue. This heating effect is absent during irradiation of tissue with a PIRL; therefore, sampling of tissues with a PIRL is a choice for purifying enzymes, because their activities are maintained.

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Language(s): eng - English
 Dates: 2019-01-062019-01-232019-03
 Publication Status: Published in print
 Pages: 7
 Publishing info: -
 Table of Contents: -
 Rev. Method: Peer
 Identifiers: DOI: 10.1021/acs.jproteome.9b00009
 Degree: -

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Project name : We acknowledge funding through the Max Planck Society in Cooperation Frauenhofer-Gesellschaft MPG-FhG “DIVE SPOT” for financial support; the research animal facilities of the Medical Center Hamburg-Eppendorf (UKE) for sample support; and Djordje Gitaric and Khashayar Shojaei-Asanjan for the design of the ablation box.
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Funding organization : -

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Title: Journal of Proteome Research
  Other : J. Proteome Res.
Source Genre: Journal
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Publ. Info: Washington, D.C. : American Chemical Society
Pages: 7 Volume / Issue: 18 (3) Sequence Number: - Start / End Page: 1451 - 1457 Identifier: ISSN: 1535-3893
CoNE: https://pure.mpg.de/cone/journals/resource/111019664290000