title: Laser ablation sample transfer for localized LC-MS/MS proteomic analysis of tissue.
abstract: We have developed a mid-infrared laser ablation sampling technique for nano-flow liquid chromatography coupled with tandem mass spectrometry proteomic profiling of discrete regions from biological samples. Laser ablation performed in transmission geometry was used to transfer material from 50-µm thick tissue sections mounted on a glass microscope slide to a capturing solvent. Captured samples were processed using filter-aided sample preparation and enzymatically digested to produce tryptic peptides for data-dependent analysis with an ion trap mass spectrometer. Comparison with ultraviolet laser capture microdissection from neighboring regions on the same tissue section revealed that infrared laser ablation transfer has higher reproducibility between samples from different consecutive sections. Both techniques allowed for proteomics investigation of different organelles without the addition of surfactants.
authors.affiliation: Department of Chemistry, Louisiana State University, Baton Rouge, Louisiana, 70803, USA.
authors.forename: Fabrizio
authors.identifier: http://orcid.org/0000-0003-1397-9919
authors.initials: F
authors.lastname: Donnarumma
authors.affiliation: Department of Chemistry, Louisiana State University, Baton Rouge, Louisiana, 70803, USA.
authors.forename: Kermit K
authors.initials: KK
authors.lastname: Murray
mesh_terms: D000818:Animals; D002853:Chromatography, Liquid / Q000379:methods*; D007259:Infrared Rays; D061346:Laser Capture Microdissection; D053685:Laser Therapy / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods; D011506:Proteins / Q000032:analysis*; D040901:Proteomics / Q000379:methods*; D051381:Rats; D013048:Specimen Handling / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D011506:Proteins
keywords: IR laser ablation; LC-MS/MS; UV laser capture microdissection; localized sampling; proteomics