title: Nanomanipulation-Coupled Matrix-Assisted Laser Desorption/ Ionization-Direct Organelle Mass Spectrometry: A Technique for the Detailed Analysis of Single Organelles.
abstract: We describe a novel technique combining precise organelle microextraction with deposition and matrix-assisted laser desorption/ionization (MALDI) for a rapid, minimally invasive mass spectrometry (MS) analysis of single organelles from living cells. A dual-positioner nanomanipulator workstation was utilized for both extraction of organelle content and precise co-deposition of analyte and matrix solution for MALDI-direct organelle mass spectrometry (DOMS) analysis. Here, the triacylglycerol (TAG) profiles of single lipid droplets from 3T3-L1 adipocytes were acquired and results validated with nanoelectrospray ionization (NSI) MS. The results demonstrate the utility of the MALDI-DOMS technique as it enabled longer mass analysis time, higher ionization efficiency, MS imaging of the co-deposited spot, and subsequent MS/MS capabilities of localized lipid content in comparison to NSI-DOMS. This method provides selective organellar resolution, which complements current biochemical analyses and prompts for subsequent subcellular studies to be performed where limited samples and analyte volume are of concern. Graphical Abstract ᅟ.
authors.affiliation: University of North Texas, Denton, TX, 76203, USA.
authors.forename: Mandy S
authors.initials: MS
authors.lastname: Phelps
authors.affiliation: University of North Texas, Denton, TX, 76203, USA.
authors.forename: Drew
authors.initials: D
authors.lastname: Sturtevant
authors.affiliation: University of North Texas, Denton, TX, 76203, USA.
authors.forename: Kent D
authors.initials: KD
authors.lastname: Chapman
authors.affiliation: University of North Texas, Denton, TX, 76203, USA. gverbeck@unt.edu.
authors.forename: Guido F
authors.initials: GF
authors.lastname: Verbeck
mesh_terms: D016475:3T3 Cells / Q000166:cytology; D000818:Animals; D004867:Equipment Design; D066292:Lipid Droplets; D059627:Liquid Phase Microextraction / Q000379:methods; D051379:Mice; D008846:Micromanipulation / Q000295:instrumentation / Q000379:methods; D015388:Organelles / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000295:instrumentation / Q000379:methods*; D053719:Tandem Mass Spectrometry
keywords: Adipocytes; Lipids; MALDI; Nanomanipulation; Organelle; Single cell