468329 In-Depth Single Cell Chemical Analysis Using Nanowell-Based Sample Preparation Combined with Ultrasensitive Capillary Electrophoresis/Mass Spectrometry

Monday, November 14, 2016: 3:15 PM
Embarcadero (Parc 55 San Francisco)
Ryan T. Kelly, Ying Zhu, Yongzheng Cong, Roza Wojcik, Ansong Charles K., Wei-Jun Qian and Richard D. Smith, Pacific Northwest National Laboratory, Richland, WA

The extension of proteomic and metabolomic analyses to the single cell level is an exciting prospect for biological research, as the intercellular heterogeneity that is currently obscured by population-level analyses can be resolved by making chemical measurements one cell at a time. Mass spectrometry (MS)-based single cell analyses require highly sensitive detection as well as essentially lossless sample preparation. We are developing a novel workflow in which cells are robotically dispensed into photolithographically patterned pico- to nanoliter wells, which are covered by an immiscible oil. Requisite sample preparation steps, including cell lysis and proteolytic digestion are performed within the nanowells. The nanowell contents are then analyzed by ultrasensitive capillary electrophoresis-mass spectrometry, or using novel gas-phase mobility based analysis technique known as structures for lossless ion manipulations (SLIM). The current state of the platform and preliminary results using mammalian cell lines will be presented.

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