Screening of Chlamydomonas reinhardtii Populations with Single-Cell Resolution by Using a High-Throughput Microscale Sample Preparation for Matrix-Assisted Laser Desorption Ionization Mass Spectrometry

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Author(s) Krismer, Jasmin, Sobek, Jens, Steinhoff, Robert F., Fagerer, Stephan R., Pabst, Martin, Zenobi, Renato
Publication Type Journal Items, Publication Status: Published
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Title Screening of Chlamydomonas reinhardtii Populations with Single-Cell Resolution by Using a High-Throughput Microscale Sample Preparation for Matrix-Assisted Laser Desorption Ionization Mass Spectrometry
Author(s) Krismer, Jasmin
Sobek, Jens
Steinhoff, Robert F.
Fagerer, Stephan R.
Pabst, Martin
Zenobi, Renato
Journal or Series Title Applied and Environmental Microbiology
Volume Number 81
Issue Number 16
Start Page 5546
End Page 5551
ISSN 0099-2240
1098-5336
Publisher American Society for Microbiology
Publication Place Washington, DC
Publication Date 2015-08
Abstract The consequences of cellular heterogeneity, such as biocide persistence, can only be tackled by studying each individual in a cell population. Fluorescent tags provide tools for the high-throughput analysis of genomes, RNA transcripts, or proteins on the single-cell level. However, the analysis of lower-molecular-weight compounds that elude tagging is still a great challenge. Here, we describe a novel high-throughput microscale sample preparation technique for single cells that allows a mass spectrum to be obtained for each individual cell within a microbial population. The approach presented includes spotting Chlamydomonas reinhardtii cells, using a noncontact microarrayer, onto a specialized slide and controlled lysis of cells separated on the slide. Throughout the sample preparation, analytes were traced and individual steps optimized using autofluorescence detection of chlorophyll. The lysates of isolated cells are subjected to a direct, label-free analysis using matrix-assisted laser desorption ionization mass spectrometry. Thus, we were able to differentiate individual cells of two Chlamydomonas reinhardtii strains based on single-cell mass spectra. Furthermore, we showed that only population profiles with real single-cell resolution render a nondistorted picture of the phenotypes contained in a population.
DOI 10.1128/AEM.01201-15
Additional Notes Published online 5 June 2015
Document Type Article
Publication Status Published
Language English
Assigned Organisational Unit(s) 02207
03430
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NEBIS System Number 000006445
Source Database ID FORM-1452524301
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@article{Krsmr2015,
  author = "Krismer, Jasmin and Sobek, Jens and Steinhoff, Robert F. and Fagerer, Stephan R. and Pabst, Martin and Zenobi, Renato",
  title = "{S}creening of {C}hlamydomonas reinhardtii {P}opulations with {S}ingle-{C}ell {R}esolution by {U}sing a {H}igh-{T}hroughput {M}icroscale {S}ample {P}reparation for {M}atrix-{A}ssisted {L}aser {D}esorption {I}onization {M}ass {S}pectrometry",
  journal = "Applied and Environmental Microbiology",
  year = 2015,
  volume = "81",
  number = "16",
  pages = "5546--5551",
  month = aug,
}


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