skip to main content
DOE PAGES title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Automated Optically Guided System for Chemical Analysis of Single Plant and Algae Cells Using Laser Microdissection/Liquid Vortex Capture/Mass Spectrometry

Abstract

Current analytical methods are not capable of providing rapid, sensitive, and comprehensive chemical analysis of a wide range of cellular constitutes of single cells (e.g., lipids, metabolites, proteins, etc.) from dispersed cell suspensions and thin tissues. This capability is important for a number of critical applications, including discovery of cellular mechanisms for coping with chemical or environmental stress and cellular response to drug treatment, to name a few. Here we introduce an optically guided platform and methodology for rapid, automated recognition, sampling, and chemical analysis of surface confined individual cells utilizing a novel hybrid laser capture microdissection/liquid vortex capture/mass spectrometry system. The system enabled automated analysis of single cells by reliably detecting and sampling them either through laser ablation from a glass microscope slide or by cutting the entire cell out of a poly(ethylene naphthalate)-coated membrane substrate that the cellular sample is deposited on. Proof of principle experiments were performed using thin tissues of Allium cepa and cultured Euglena gracilis and Phacus cell suspensions as model systems for single cell analysis using the developed method. Reliable, hands-off laser ablation sampling coupled to liquid vortex capture/mass spectrometry analysis was conducted for hundreds of individual Allium cepa cells in connected tissue. Inmore » addition, more than 300 individual Euglena gracilis and Phacus cells were analyzed automatically and sampled using laser microdissection sampling with the same liquid vortex capture/mass spectrometry analysis system. Principal component analysis-linear discriminant analysis, applied to each mass spectral dataset, was used to determine the accuracy of differentiation of the different algae cell lines.« less

Authors:
;
Publication Date:
Research Org.:
Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22)
OSTI Identifier:
1465213
Alternate Identifier(s):
OSTI ID: 1559740
Grant/Contract Number:  
AC05-00OR22725
Resource Type:
Published Article
Journal Name:
Frontiers in Plant Science
Additional Journal Information:
Journal Name: Frontiers in Plant Science Journal Volume: 9; Journal ID: ISSN 1664-462X
Publisher:
Frontiers Research Foundation
Country of Publication:
Switzerland
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Cahill, John F., and Kertesz, Vilmos. Automated Optically Guided System for Chemical Analysis of Single Plant and Algae Cells Using Laser Microdissection/Liquid Vortex Capture/Mass Spectrometry. Switzerland: N. p., 2018. Web. doi:10.3389/fpls.2018.01211.
Cahill, John F., & Kertesz, Vilmos. Automated Optically Guided System for Chemical Analysis of Single Plant and Algae Cells Using Laser Microdissection/Liquid Vortex Capture/Mass Spectrometry. Switzerland. doi:10.3389/fpls.2018.01211.
Cahill, John F., and Kertesz, Vilmos. Mon . "Automated Optically Guided System for Chemical Analysis of Single Plant and Algae Cells Using Laser Microdissection/Liquid Vortex Capture/Mass Spectrometry". Switzerland. doi:10.3389/fpls.2018.01211.
@article{osti_1465213,
title = {Automated Optically Guided System for Chemical Analysis of Single Plant and Algae Cells Using Laser Microdissection/Liquid Vortex Capture/Mass Spectrometry},
author = {Cahill, John F. and Kertesz, Vilmos},
abstractNote = {Current analytical methods are not capable of providing rapid, sensitive, and comprehensive chemical analysis of a wide range of cellular constitutes of single cells (e.g., lipids, metabolites, proteins, etc.) from dispersed cell suspensions and thin tissues. This capability is important for a number of critical applications, including discovery of cellular mechanisms for coping with chemical or environmental stress and cellular response to drug treatment, to name a few. Here we introduce an optically guided platform and methodology for rapid, automated recognition, sampling, and chemical analysis of surface confined individual cells utilizing a novel hybrid laser capture microdissection/liquid vortex capture/mass spectrometry system. The system enabled automated analysis of single cells by reliably detecting and sampling them either through laser ablation from a glass microscope slide or by cutting the entire cell out of a poly(ethylene naphthalate)-coated membrane substrate that the cellular sample is deposited on. Proof of principle experiments were performed using thin tissues of Allium cepa and cultured Euglena gracilis and Phacus cell suspensions as model systems for single cell analysis using the developed method. Reliable, hands-off laser ablation sampling coupled to liquid vortex capture/mass spectrometry analysis was conducted for hundreds of individual Allium cepa cells in connected tissue. In addition, more than 300 individual Euglena gracilis and Phacus cells were analyzed automatically and sampled using laser microdissection sampling with the same liquid vortex capture/mass spectrometry analysis system. Principal component analysis-linear discriminant analysis, applied to each mass spectral dataset, was used to determine the accuracy of differentiation of the different algae cell lines.},
doi = {10.3389/fpls.2018.01211},
journal = {Frontiers in Plant Science},
number = ,
volume = 9,
place = {Switzerland},
year = {2018},
month = {8}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record
DOI: 10.3389/fpls.2018.01211

Citation Metrics:
Cited by: 1 work
Citation information provided by
Web of Science

Save / Share:

Works referenced in this record:

Autonomous Metabolic Oscillations Robustly Gate the Early and Late Cell Cycle
journal, January 2017


In Situ Metabolic Profiling of Single Cells by Laser Ablation Electrospray Ionization Mass Spectrometry
journal, October 2009

  • Shrestha, Bindesh; Vertes, Akos
  • Analytical Chemistry, Vol. 81, Issue 20
  • DOI: 10.1021/ac901525g

Laser dissection sampling modes for direct mass spectral analysis: Laser dissection sampling modes for mass spectral analysis
journal, February 2016

  • Cahill, John F.; Kertesz, Vilmos; Van Berkel, Gary J.
  • Rapid Communications in Mass Spectrometry, Vol. 30, Issue 5
  • DOI: 10.1002/rcm.7477

Single Cell Peptide Heterogeneity of Rat Islets of Langerhans
journal, July 2016


UPLC-MS analysis of Chlamydomonas reinhardtii and Scenedesmus obliquus lipid extracts and their possible metabolic roles
journal, September 2014

  • Sharma, Durlubh Kumar; Gautam, Kshipra; Jueppner, Jessica
  • Journal of Applied Phycology, Vol. 27, Issue 3
  • DOI: 10.1007/s10811-014-0407-2

Single Cell Analysis Reveals the Stochastic Phase of Reprogramming to Pluripotency Is an Ordered Probabilistic Process
journal, April 2014


Intelligent image-based in situ single-cell isolation
journal, January 2018


Laser microdissection and atmospheric pressure chemical ionization mass spectrometry coupled for multimodal imaging: LA-APCI/MS imaging
journal, May 2013

  • Lorenz, Matthias; Ovchinnikova, Olga S.; Kertesz, Vilmos
  • Rapid Communications in Mass Spectrometry, Vol. 27, Issue 13
  • DOI: 10.1002/rcm.6593

Direct imaging of single cells and tissue at sub-cellular spatial resolution using transmission geometry MALDI MS
journal, November 2012

  • Zavalin, Andre; Todd, Erik M.; Rawhouser, Patrick D.
  • Journal of Mass Spectrometry, Vol. 47, Issue 11
  • DOI: 10.1002/jms.3132

Categorizing Cells on the Basis of their Chemical Profiles: Progress in Single-Cell Mass Spectrometry
journal, February 2017

  • Comi, Troy J.; Do, Thanh D.; Rubakhin, Stanislav S.
  • Journal of the American Chemical Society, Vol. 139, Issue 11
  • DOI: 10.1021/jacs.6b12822

Live single-cell video-mass spectrometry for cellular and subcellular molecular detection and cell classification
journal, December 2008

  • Mizuno, Hajime; Tsuyama, Naohiro; Harada, Takanori
  • Journal of Mass Spectrometry, Vol. 43, Issue 12
  • DOI: 10.1002/jms.1460

High-density micro-arrays for mass spectrometry
journal, January 2010

  • Urban, Pawel L.; Jefimovs, Konstantins; Amantonico, Andrea
  • Lab on a Chip, Vol. 10, Issue 23
  • DOI: 10.1039/c0lc00211a

Solvent effects on differentiation of mouse brain tissue using laser microdissection ‘cut and drop’ sampling with direct mass spectral analysis
journal, February 2018

  • Cahill, John F.; Kertesz, Vilmos; Porta, Tiffany
  • Rapid Communications in Mass Spectrometry, Vol. 32, Issue 5
  • DOI: 10.1002/rcm.8053

Single-Cell Metabolomics: Analytical and Biological Perspectives
journal, December 2013


Mass Cytometry: Single Cells, Many Features
journal, May 2016


Single-cell MALDI: a new tool for direct peptide profiling
journal, April 2000

  • Li, Lingjun; Garden, Rebecca W.; Sweedler, Jonathan V.
  • Trends in Biotechnology, Vol. 18, Issue 4, p. 151-160
  • DOI: 10.1016/S0167-7799(00)01427-X