Laser Capture Microdissection–Liquid Vortex Capture Mass Spectrometry Metabolic Profiling of Single Onion Epidermis and Microalgae Cells
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Laser capture microdissection is a valuable technique in individually isolating single cells whether in tissue networks or deposited from a cell suspension. New developments have enabled coupling of laser capture microdissection with mass spectrometry via liquid vortex capture sampling probe. This enables online metabolic profiling of sectioned cells. Here, we describe the protocol used to deposit, isolate, and individually chemically characterize single Allium cepa and Chlamydomonas reinhardtii cells by laser capture microdissection–liquid vortex capture mass spectrometry.
Key wordsLaser capture microdissection Mass spectrometry Single cell Onion epidermis cell Microalgae Liquid-vortex capture Metabolites
This work and J. F. C. was supported by the U.S. Department of Energy, Office of Science, Basic Energy Sciences, Chemical Sciences, Geosciences, and Biosciences Division and the Laboratory Directed Research and Development Program of Oak Ridge National Laboratory, managed by UT-Battelle, LLC, for the U.S. Department of Energy. V. K. was supported by the U. S. Department of Energy, Office of Science, Basic Energy Sciences, Chemical Sciences, Geosciences, and Biosciences Division.
- 7.Shrestha B, Vertes A (2010) Direct analysis of single cells by mass spectrometry at atmospheric pressure. J Vis Exp (43):e2144. https://doi.org/10.3791/2144
- 8.Cahill JF, Kertesz V, Van Berkel GJ (2015) Characterization and application of a hybrid optical microscopy/laser ablation liquid vortex capture/electrospray ionization system for mass spectrometry imaging with sub-micrometer spatial resolution. Anal Chem 87(21):11113–11121. https://doi.org/10.1021/acs.analchem.5b03293CrossRefGoogle Scholar
- 9.Cahill JF, Kertesz V, Porta T, LeBlanc JCY, Heeren RMA, Van Berkel GJ (2018) Solvent effects on differentiation of mouse brain tissue using laser microdissection ‘cut and drop’ sampling with direct mass spectral analysis. Rapid Commun Mass Spectrom 32(5):414–422. https://doi.org/10.1002/rcm.8053CrossRefPubMedGoogle Scholar
- 10.Cahill JF, Kertesz V, Weiskittel TM, Vavrek M, Freddo C, Van Berkel GJ (2016) Online, absolute quantitation of propranolol from spatially distinct 20-and 40-μm dissections of brain, liver, and kidney thin tissue sections by laser microdissection-liquid vortex capture-mass spectrometry. Anal Chem 88(11):6026–6034. https://doi.org/10.1021/acs.analchem.6b01155CrossRefPubMedGoogle Scholar