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Profiling and quantifying endogenous molecules in single cells using nano-DESI MS
Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Kemiska sektionen, Institutionen för kemi - BMC, Analytisk kemi. (Lanekoff)
Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Kemiska sektionen, Institutionen för kemi - BMC, Analytisk kemi. (Lanekoff)
2017 (Engelska)Ingår i: The Analyst, ISSN 0003-2654, E-ISSN 1364-5528, Vol. 142, nr 19, s. 3639-3647Artikel i tidskrift (Refereegranskat) Published
Abstract [en]

Molecular profiling of single cells has the potential to significantly advance our understanding of cell function and cellular processes of importance to health and disease. In particular, small molecules with rapid turn-over rates can reveal activated metabolic pathways resulting from an altered chemical environment or cellular events such as differentiation. Consequently, techniques for quantitative metabolite detection acquired in a higher throughput manner are needed to characterize the biological variability between seemingly homogenous cells. Here, we show that nanospray desorption electrospray ionization (nano-DESI) mass spectrometry ( MS) enables sensitive molecular profiling and quantification of endogenous species in single cells in a higher throughput manner. Specifically, we show a large number of detected amino acids and phospholipids, including plasmalogens, readily detected from single cheek cells. Further, by incorporating a phosphatidylcholine ( PC) internal standard into the nano-DESI solvent, we determined the total amount of PC in one cell to be 1.2 pmoles. Finally, we describe a higher throughput approach where molecules in single cells are automatically profiled. These developments in single cell analysis provide a basis for future studies to understand cellular processes related to drug effects, cell differentiation and altered chemical microenvironments.

Ort, förlag, år, upplaga, sidor
ROYAL SOC CHEMISTRY , 2017. Vol. 142, nr 19, s. 3639-3647
Nationell ämneskategori
Analytisk kemi
Forskningsämne
Kemi med inriktning mot analytisk kemi
Identifikatorer
URN: urn:nbn:se:uu:diva-336430DOI: 10.1039/c7an00885fISI: 000411703800013PubMedID: 28835951OAI: oai:DiVA.org:uu-336430DiVA, id: diva2:1166526
Forskningsfinansiär
Vetenskapsrådet, VR 621-2013-4231Stiftelsen för strategisk forskning (SSF), SSF ICA-6Tillgänglig från: 2017-12-15 Skapad: 2017-12-15 Senast uppdaterad: 2018-11-29
Ingår i avhandling
1. Applications of nanospray desorption electrospray ionization mass spectrometry: In situ lipid and metabolite analysis from cells to tissue
Öppna denna publikation i ny flik eller fönster >>Applications of nanospray desorption electrospray ionization mass spectrometry: In situ lipid and metabolite analysis from cells to tissue
2018 (Engelska)Doktorsavhandling, sammanläggning (Övrigt vetenskapligt)
Abstract [en]

Ambient mass spectrometry (MS) has proved to be an important addition to the bioanalytical toolbox. These methods perform analyte sampling and ionization under atmospheric pressure, and require very little sample preparation other than the sampling process in front of the machine. Nanospray desorption electrospray ionization (nano-DESI) is an ambient MS technique developed in 2010 that utilizes localized liquid extraction for surface sampling. The aim of this thesis was to explore the possibilities of this technique, and identify areas in which nano-DESI MS could further contribute to the community of MS-based surface analysis.

One such area was found to be mass spectrometry imaging (MSI) of small-molecule neurotransmitters. By the use of deuterated standards of acetylcholine, γ-aminobutyric acid and glutamate, the respective endogenous compounds were successfully imaged in coronal sections of rat brain. The use of internal standards was shown to be essential to compensatee for matrix effects in different regions of the brain. In a second imaging study, nano-DESI MSI was used to compare the chemical profiles of diabetic rat kidney tissue and control. Analysis was performed on kidney two weeks after diabetic onset, before any pathohistological changes relating to diabetic nephropathy can be seen in a microscope. In our study, it was shown that a large number of chemical species related to energy metabolism were detected with altered signal intensity in diabetic kidney tissue.

To push the limits of nano-DESI analysis, its use for single-cell analysis was evaluated. By placing buccal epithelial cells in contact with the nano-DESI probe, it was possible to identify 46 endogenous compounds and detect differences between cells from three human donors. In addition, it was shown that molecules from single cells on a surface could be detected by scanning the surface with the nano-DESI probe, which opens up for development of an automated analysis with higher throughput.

The last study in this thesis was concerned with method development rather than application, as it presented a setup for pneumatically assisted nano-DESI. Evaluation showed that the setup provided improved sensitivity in the analysis of small metabolites, and provided the possibility of using pure water as nano-DESI solvent.

Ort, förlag, år, upplaga, sidor
Uppsala: Acta Universitatis Upsaliensis, 2018. s. 78
Serie
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology, ISSN 1651-6214 ; 1660
Nyckelord
Mass spectrometry, mass spectrometry imaging (MSI), nanospray desorption electrospray ionization (nano-DESI), single-cell analysis, neurotransmitter imaging, diabetic nephropathy, pneumatic nebulization, lipidomics, metabolomics
Nationell ämneskategori
Kemi
Forskningsämne
Kemi med inriktning mot analytisk kemi
Identifikatorer
urn:nbn:se:uu:diva-347674 (URN)978-91-513-0307-9 (ISBN)
Disputation
2018-05-25, A1:107a, BMC, Husargatan 3, Uppsala, 13:00 (Engelska)
Opponent
Handledare
Tillgänglig från: 2018-05-03 Skapad: 2018-04-05 Senast uppdaterad: 2018-10-08

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Bergman, Hilde-MarleneLanekoff, Ingela

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