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Direct Analysis of Single Cells by Mass Spectrometry at Atmospheric Pressure
Analysis of biochemicals in single cells is important for understanding cell metabolism, cell cycle, adaptation, disease states, etc. Even the same cell types exhibit heterogeneous biochemical makeup depending on their physiological conditions and interactions with the environment. Conventional meth...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MyJove Corporation
2010
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3157873/ https://www.ncbi.nlm.nih.gov/pubmed/20834224 http://dx.doi.org/10.3791/2144 |
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author | Shrestha, Bindesh Vertes, Akos |
author_facet | Shrestha, Bindesh Vertes, Akos |
author_sort | Shrestha, Bindesh |
collection | PubMed |
description | Analysis of biochemicals in single cells is important for understanding cell metabolism, cell cycle, adaptation, disease states, etc. Even the same cell types exhibit heterogeneous biochemical makeup depending on their physiological conditions and interactions with the environment. Conventional methods of mass spectrometry (MS) used for the analysis of biomolecules in single cells rely on extensive sample preparation. Removing the cells from their natural environment and extensive sample processing could lead to changes in the cellular composition. Ambient ionization methods enable the analysis of samples in their native environment and without extensive sample preparation.(1) The techniques based on the mid infrared (mid-IR) laser ablation of biological materials at 2.94 μm wavelength utilize the sudden excitation of water that results in phase explosion.(2) Ambient ionization techniques based on mid-IR laser radiation, such as laser ablation electrospray ionization (LAESI) and atmospheric pressure infrared matrix-assisted laser desorption ionization (AP IR-MALDI), have successfully demonstrated the ability to directly analyze water-rich tissues and biofluids at atmospheric pressure.(3-11) In LAESI the mid-IR laser ablation plume that mostly consists of neutral particulate matter from the sample coalesces with highly charged electrospray droplets to produce ions. Recently, mid-IR ablation of single cells was performed by delivering the mid-IR radiation through an etched fiber. The plume generated from this ablation was postionized by an electrospray enabling the analysis of diverse metabolites in single cells by LAESI-MS.(12) This article describes the detailed protocol for single cell analysis using LAESI-MS. The presented video demonstrates the analysis of a single epidermal cell from the skin of an Allium cepa bulb. The schematic of the system is shown in Figure 1. A representative example of single cell ablation and a LAESI mass spectrum from the cell are provided in Figure 2. |
format | Online Article Text |
id | pubmed-3157873 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2010 |
publisher | MyJove Corporation |
record_format | MEDLINE/PubMed |
spelling | pubmed-31578732011-11-15 Direct Analysis of Single Cells by Mass Spectrometry at Atmospheric Pressure Shrestha, Bindesh Vertes, Akos J Vis Exp Cellular Biology Analysis of biochemicals in single cells is important for understanding cell metabolism, cell cycle, adaptation, disease states, etc. Even the same cell types exhibit heterogeneous biochemical makeup depending on their physiological conditions and interactions with the environment. Conventional methods of mass spectrometry (MS) used for the analysis of biomolecules in single cells rely on extensive sample preparation. Removing the cells from their natural environment and extensive sample processing could lead to changes in the cellular composition. Ambient ionization methods enable the analysis of samples in their native environment and without extensive sample preparation.(1) The techniques based on the mid infrared (mid-IR) laser ablation of biological materials at 2.94 μm wavelength utilize the sudden excitation of water that results in phase explosion.(2) Ambient ionization techniques based on mid-IR laser radiation, such as laser ablation electrospray ionization (LAESI) and atmospheric pressure infrared matrix-assisted laser desorption ionization (AP IR-MALDI), have successfully demonstrated the ability to directly analyze water-rich tissues and biofluids at atmospheric pressure.(3-11) In LAESI the mid-IR laser ablation plume that mostly consists of neutral particulate matter from the sample coalesces with highly charged electrospray droplets to produce ions. Recently, mid-IR ablation of single cells was performed by delivering the mid-IR radiation through an etched fiber. The plume generated from this ablation was postionized by an electrospray enabling the analysis of diverse metabolites in single cells by LAESI-MS.(12) This article describes the detailed protocol for single cell analysis using LAESI-MS. The presented video demonstrates the analysis of a single epidermal cell from the skin of an Allium cepa bulb. The schematic of the system is shown in Figure 1. A representative example of single cell ablation and a LAESI mass spectrum from the cell are provided in Figure 2. MyJove Corporation 2010-09-04 /pmc/articles/PMC3157873/ /pubmed/20834224 http://dx.doi.org/10.3791/2144 Text en Copyright © 2010, Journal of Visualized Experiments http://creativecommons.org/licenses/by-nc-nd/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License. To view a copy of this license, visithttp://creativecommons.org/licenses/by-nc-nd/3.0/ |
spellingShingle | Cellular Biology Shrestha, Bindesh Vertes, Akos Direct Analysis of Single Cells by Mass Spectrometry at Atmospheric Pressure |
title | Direct Analysis of Single Cells by Mass Spectrometry at Atmospheric Pressure |
title_full | Direct Analysis of Single Cells by Mass Spectrometry at Atmospheric Pressure |
title_fullStr | Direct Analysis of Single Cells by Mass Spectrometry at Atmospheric Pressure |
title_full_unstemmed | Direct Analysis of Single Cells by Mass Spectrometry at Atmospheric Pressure |
title_short | Direct Analysis of Single Cells by Mass Spectrometry at Atmospheric Pressure |
title_sort | direct analysis of single cells by mass spectrometry at atmospheric pressure |
topic | Cellular Biology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3157873/ https://www.ncbi.nlm.nih.gov/pubmed/20834224 http://dx.doi.org/10.3791/2144 |
work_keys_str_mv | AT shresthabindesh directanalysisofsinglecellsbymassspectrometryatatmosphericpressure AT vertesakos directanalysisofsinglecellsbymassspectrometryatatmosphericpressure |