TY - JOUR
T1 - 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
AU - Krismer, Jasmin
AU - Sobek, Jens
AU - Steinhoff, Robert F.
AU - Fagerer, Stephan R.
AU - Pabst, Martin
AU - Zenobi, Renato
PY - 2015
Y1 - 2015
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=84937964198&partnerID=8YFLogxK
U2 - 10.1128/AEM.01201-15
DO - 10.1128/AEM.01201-15
M3 - Article
C2 - 26048935
AN - SCOPUS:84937964198
SN - 0099-2240
VL - 81
SP - 5546
EP - 5551
JO - Applied and Environmental Microbiology
JF - Applied and Environmental Microbiology
IS - 16
ER -