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Nanoliter-scale Autosampler for Seamless Integration of NanoPOTS Platform with LC-MS

EMSL Project ID


We propose to develop a high throughput, automated, and ultrasensitive single cell proteomic platform by seamlessly integrating our nanodroplet Processing in One-pot for Trace Sample technique with ultrasensitive liquid chromatography mass spectrometry. To accomplish our goal, the following Specific Aims will be pursued:
Aims 1: We will develop a new microfluidic sampling probe to directly introduce nanodroplets with volumes < 200 nL into high pressure valves.
Aims 2: We will develop an integrated nanoliter autosampler with analysis throughput of >50 samples per day.
Aims 3: We will demonstrate the utility of the new platform in proteomics-based typing of plant cells isolated from Brachypodium.

Project Details

Start Date
End Date


Principal Investigator

Ying Zhu
Environmental Molecular Sciences Laboratory

Team Members

Andrey Liyu
Environmental Molecular Sciences Laboratory

Daniel Orton
Pacific Northwest National Laboratory

Ronald Moore
Pacific Northwest National Laboratory

Related Publications

Dou M., C. Tsai, P.D. Piehowski, Y. Wang, T.L. Fillmore, R. Zhao, and R.J. Moore, et al. 2019. "Automated Nanoflow Two-Dimensional Reversed-Phase Liquid Chromatography System Enables In-Depth Proteome and Phosphoproteome Profiling of Nanoscale Samples." Analytical Chemistry 91, no. 15:9707-9715. PNNL-SA-141437. doi:10.1021/acs.analchem.9b01248
Dou M., G. Clair, C. Tsai, K. Xu, W.B. Chrisler, R.L. Sontag, and R. Zhao, et al. 2019. "High-Throughput Single Cell Proteomics Enabled by Multiplex Isobaric Labelling in a Nanodroplet Sample Preparation Platform." Analytical Chemistry 91, no. 20:13119-13127. PNNL-SA-146297. doi:10.1021/acs.analchem.9b03349
Liu W wen, Zhu Y (2020) ?Development and application of analytical detection techniques for droplet-based microfluidics?-A review. Anal Chim Acta 1113: 66?84.
Williams SM, Liyu A V., Tsai CF, Moore RJ, Orton DJ, Chrisler WB, Gaffrey MJ, Liu T, Smith RD, Kelly RT, et al. (2020) Automated Coupling of Nanodroplet Sample Preparation with Liquid Chromatography-Mass Spectrometry for High-Throughput Single-Cell Proteomics. Anal Chem 92: 10588?10596.
Xiang P., Y. Zhu, Y. Yang, Z. Zhao, S.M. Williams, R.J. Moore, and R.T. Kelly, et al. 2020. "Picoflow Liquid Chromatography-Mass Spectrometry for Ultrasensitive Bottom-up Proteomics using 2-µm i.d. Open Tubular Columns." Analytical Chemistry 92, no. 7:4711-4715. PNNL-SA-150120. doi:10.1021/acs.analchem.9b05639
Zhu Y, Scheibinger M, Ellwanger DC, Krey JF, Choi D, Kelly RT, Heller S, Barr-Gillespie PG (2019) Single-cell proteomics reveals changes in expression during hair-cell development. Elife 8: e50777.