Sample preparation for single-cell MS analysis | Edward Emmott | SCP2019

A major limitation to applying quantitative LC-MS/MS proteomics to small samples, such as single cells, are the losses incurred during sample cleanup. We discuss our revised SCoPE2 pipeline for processing single-cell samples, building on the SCoPE-MS method using mPOP lysis (Specht et al. 2018, bioRxiv). This involves using only mass spectrometry-compatible reagents, as well as a move to freeze-heat lysis of samples. This removes the need for sample cleanup and permits automation and higher throughput sample processing. We will explain how proceed from FACS-sorted single cells in multi-well plates to semi-automated SCoPE2 sample preparation.

Relevant publications for further reading:

Specht H, Emmott E, Koller T, Slavov N (2019)

Specht H, Harmange G, Perlman DH, Emmott E, Niziolek Z, Budnik B, Slavov N. (2018)
Automated sample preparation for high-throughput single-cell proteomics

Huffman RG, Specht H, Chen AT, Slavov N. (2019)
DO-MS: Data-Driven Optimization of Mass Spectrometry Methods

Chen AT, Franks A, Slavov N. (2019)
DART-ID increases single-cell proteome coverage
Specht H, and Slavov N. (2018)
Transformative opportunities for single cell proteomics

Budnik B., Levy E., Harmange G., Slavov N. (2018)
SCoPE-MS: mass-spectrometry of single mammalian cells quantifies proteome heterogeneity during cell differentiation

[Hwafa]

Such a clear and well explained presentation ! So happy I could attend to SCP 2020 this summer!

[桓林-gv]

May I ask a question? Cells were sorted in 1uL of 25fmol or 25pmol Waters Massprep? Your description was different between the beginning and summary part 1.