Abstract by Yiran Liang
Chemistry and Biochemistry
A low-cost pipetting platform for automated preparation of nanoscale and single-cell proteomic samples
Proteomic analyses are often limited by the amount of available starting materials. Despite the high performence of our nanoPOTS platform, the dissemination of nanoPOTS to other laboratories has been limited due to its cost and complexity and the required engineering and microfluidic expertise. Here we describe a low-cost, fully automated workflow using the Opentrons 2 (OT-2) robot and an autosampler to prepare samples comprising 1–500 cells. The unmodified OT-2 liquid handler was used to dispense 1–10 µL aqueous solutions with CV values of <5.8% and dispensing accuracy <7%. The evaporation rate in the well plate was calculated and a program was developed to periodically dispense buffer or water into the wells. An UltiMate 3000 autosampler (Thermo) was modified with a nano-volume 10-port for automated and ultrasensitive analysis of the prepared samples. Using a 30 µm column, ~150 protein groups and 800 unique peptides were identified from one Hela cell. Thus the low cost microliter liquid handler offers strong potential to broadly increase accessibility of single cell proteomic analyses to other laboratories.