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Single Cell Metabolism pp 159–179Cite as

Ambient Lipidomic Analysis of Single Mammalian Oocytes and Preimplantation Embryos Using Desorption Electrospray Ionization (DESI) Mass Spectrometry

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 2064))

Abstract

Desorption electrospray ionization (DESI) is a spray-based ambient ionization method for mass spectrometry (MS) that generates ions in native atmospheric conditions (e.g., pressure and temperature). Ambient ionization allows in situ analysis of unmodified samples by eliminating analyte extraction and separation steps before MS. Lipid analysis of individual mammalian oocytes and preimplantation embryos is challenging because of their complex chemical composition and minute dimensions (100–300 μm in diameter). Nonetheless, DESI-MS can provide comprehensive lipidomic profiles of individual oocytes or embryos using a fast and simple workflow. DESI-MS lipid profiles include many classes of lipids such as phosphatidylcholines (PC), triacylglycerols (TAG), free fatty acids (FFA), phosphatidylethanolamines (PE), phosphatidylinositols (PI), phosphatidylserines (PS), diacylglycerols (DAG), ubiquinone, cholesterol and cholesterol derivatives (e.g., cholesterol sulfate and cholesterol esters). Depending on the mass spectrometer used, there is the additional possibility of obtaining structural information of lipids via MSn, and molecular formulae via high resolution MS. Here, we describe the procedures for sample handling, the DESI-MS experimental arrangement, and the data collection and data analysis using low and high-resolution mass spectrometers (such as a linear ion trap and Orbitrap mass analyzer, respectively), as well as strategies for structural characterization of lipids. Lastly, we detail our workflow for relating the chemical information obtained by DESI-MS into the biological context of oocyte and embryo metabolism.

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Acknowledgments

This work was supported by NSF CHE 1307264. C.R.F. was supported from the Purdue University Center for Cancer Research Small Grants and from the Brazilian National Council for Scientific and Technological Development (CNPq). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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Authors and Affiliations

  1. Department of Chemistry and Center for Analytical Instrumentation Development, Purdue University, West Lafayette, IN, USA

    Christina R. Ferreira, Valentina Pirro, Alan K. Jarmusch, Clint M. Alfaro & R. Graham Cooks

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  1. Christina R. Ferreira
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  2. Valentina Pirro
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  3. Alan K. Jarmusch
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  4. Clint M. Alfaro
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  5. R. Graham Cooks
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Corresponding author

Correspondence to Christina R. Ferreira .

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  1. Waters Corporation, Milford, MA, USA

    Bindesh Shrestha

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Ferreira, C.R., Pirro, V., Jarmusch, A.K., Alfaro, C.M., Cooks, R.G. (2020). Ambient Lipidomic Analysis of Single Mammalian Oocytes and Preimplantation Embryos Using Desorption Electrospray Ionization (DESI) Mass Spectrometry. In: Shrestha, B. (eds) Single Cell Metabolism. Methods in Molecular Biology, vol 2064. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9831-9_13

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  • DOI: https://doi.org/10.1007/978-1-4939-9831-9_13

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9829-6

  • Online ISBN: 978-1-4939-9831-9

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