Dataset: Mass spectrometry analysis of single mammalian cells obtained using the redesigned T-probe from laboratory experiments performed in 2017 and 2018

ValidatedFinal no updates expectedDOI: 10.26008/1912/bco-dmo.851838.1Version 1 (2021-05-11)Dataset Type:experimental

Principal Investigator: Zhibo Yang (University of Oklahoma)

Co-Principal Investigator: Linda Atkinson (University of Oklahoma)

BCO-DMO Data Manager: Amber D. York (Woods Hole Oceanographic Institution)


Project: Collaborative Research: Creatine Cycling in Marine Bacterial and Phytoplankton Assemblages (Creatine Cycling)


Abstract

Mass spectrometry analysis of single mammalian cells obtained using the redesigned T-probe from laboratory experiments performed in 2017 and 2018. These results were published in Zhu et al. (2019).

The following are excerpts from Zhu et al., 2019. Please refer to this publication for more details.

Methodology

During the SCMS analysis, the redesigned T-probe was coupled to the in-home developed SCMS platform employed in our previous SCMS studies. Briefly, this platform includes an XYZ-translational stage system, two digital microscopes, and a Thermo LTQ Orbitrap XL mass spectrometer. Cells in both control and drug treatment groups were used for the SCMS experiments (detailed sample preparation procedures are described in the Supporting Information). Irinotecan is a common anticancer drug for the treatment of colon cancer that inhibits the function of Topoisomerase I, leading to DNA damage and cell apoptosis. This drug compound was selected to treat live HCT-116 colorectal cells in our experiments to demonstrate the change of cellular metabolites upon the treatment of anticancer agent. Specifically, cells were first treated using 18 mM irinotecan for 45 min, and then rinsed and detached using trypsinization. Afterwards, a droplet of cell suspension solution was placed onto a glass slide, which was attached to the XYZ-stage system controlled by a LabView software package (incremental step size = 0.1 mm). Using two digital microscopes as the visual guide, the sampling probe tip initially located above the sample plate was submerged into the solution containing cells by lifting the Z-stage. Upon selecting a target cell, the sampling probe can precisely draw the target cell with visual guidance. The XYZ-stage was then immediately lowered down to free the sampling probe tip from the culture medium and stop the suction of culture medium. Due to the complex composition of the cell culture medium that may affect the detection sensitivity, caution should be taken to minimize its amount withdrawn during cell sampling. This is particularly important for future analysis of patient cells suspended in complex biological fluids such as blood, urine, saliva, and cerebrospinal fluid (CSF). After a single cell was withdrawn, the solvent provided through the solvent-providing capillary (flowrate = 0.5 mL/min) mixed with the cell at the T-junction, and cell lysis rapidly occurred inside the nano-ESI emitter. In our SCMS analysis, an ionization voltage (~4 kV) was applied to the conductive union and transmitted throughout the solution inside the solvent-providing capillary and the nano-ESI emitter to ionize the cell lysis for MS analysis.

Instrument

Thermo LTQ Orbitrap XL mass spectrometer (Thermo Scientific, Waltham, MA, United States). Mass analyze parameters were as follows: mass resolution 60,000, +4 kV ionization voltage at positive ion mode (0.05–0.07 μA of ion current), 1 microscan, 100 ms max injection time, and automatic gain control on.

Location of experiments: University of Oklahoma, Norman, OK 73019
Cell line: HCT-116 (human colon cancer cell line)


Related Datasets

No Related Datasets

Related Publications

Results

Zhu, Y., Liu, R., & Yang, Z. (2019). Redesigning the T-probe for mass spectrometry analysis of online lysis of non-adherent single cells. Analytica Chimica Acta, 1084, 53–59. doi:10.1016/j.aca.2019.07.059
Methods

Liu, R., Pan, N., Zhu, Y., & Yang, Z. (2018). T-Probe: An Integrated Microscale Device for Online In Situ Single Cell Analysis and Metabolic Profiling Using Mass Spectrometry. Analytical Chemistry, 90(18), 11078–11085. doi:10.1021/acs.analchem.8b02927
Methods

Pan, N., Rao, W., Kothapalli, N. R., Liu, R., Burgett, A. W. G., & Yang, Z. (2014). The Single-Probe: A Miniaturized Multifunctional Device for Single Cell Mass Spectrometry Analysis. Analytical Chemistry, 86(19), 9376–9380. doi:10.1021/ac5029038
Methods

Pan, N., Rao, W., Standke, S. J., & Yang, Z. (2016). Using Dicationic Ion-Pairing Compounds To Enhance the Single Cell Mass Spectrometry Analysis Using the Single-Probe: A Microscale Sampling and Ionization Device. Analytical Chemistry, 88(13), 6812–6819. doi:10.1021/acs.analchem.6b01284
Methods

Rao, W., Pan, N., & Yang, Z. (2016). Applications of the Single-probe: Mass Spectrometry Imaging and Single Cell Analysis under Ambient Conditions. Journal of Visualized Experiments, (112). doi:10.3791/53911