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Online Analysis of Single Cyanobacteria and Algae Cells under Nitrogen-Limited Conditions Using Aerosol Time-of-Flight Mass Spectrometry
journal contribution
posted on 2015-08-18, 00:00 authored by John F. Cahill, Thomas K. Darlington, Christine Fitzgerald, Nathan G. Schoepp, Joris Beld, Michael
D. Burkart, Kimberly A. PratherMetabolomics studies typically perform
measurements on populations
of whole cells which provide the average representation of a collection
of many cells. However, key mechanistic information can be lost using
this approach. Investigating chemistry at the single cell level yields
a more accurate representation of the diversity of populations within
a cell sample; however, this approach has many analytical challenges.
In this study, an aerosol time-of-flight mass spectrometer (ATOFMS)
was used for rapid analysis of single algae and cyanobacteria cells
with diameters ranging from 1 to 8 μm. Cells were aerosolized
by nebulization and directly transmitted into the ATOFMS. Whole cells
were determined to remain intact inside the instrument through a combination
of particle sizing and imaging measurements. Differences in cell populations
were observed after perturbing Chlamydomonas reinhardtii cells via nitrogen deprivation. Thousands of single cells were measured
over a period of 4 days for nitrogen-replete and nitrogen-limited
conditions. A comparison of the single cell mass spectra of the cells
sampled under the two conditions revealed an increase in the dipalmitic
acid sulfolipid sulfoquinovosyldiacylglycerol (SQDG), a chloroplast
membrane lipid, under nitrogen-limited conditions. Single cell peak
intensity distributions demonstrate the ability of the ATOFMS to measure
metabolic differences of single cells. The ATOFMS provides an unprecedented
maximum throughput of 50 Hz, enabling the rapid online measurement
of thousands of single cell mass spectra.
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Single cell peak intensity distributionscell populationscell mass spectraSQDGnitrogen deprivation50 Hzimaging measurements8 μ mAlgae CellsATOFMScell level yieldsWhole cellsSingle Cyanobacteria4 dayschloroplast membrane lipidOnline Analysiscyanobacteria cellsparticle sizingcell sampledipalmitic acid sulfolipid sulfoquinovosyldiacylglycerolperturbing Chlamydomonas reinhardtii cells
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