MassIVE MSV000093330

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Multimodal measurement of the transcriptome and proteome from dissociated human pancreatic islet cells using nanoSPLITS

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Description

Human pancreatic islets from two human donors (Prodo Laboratories) were acquired and cultured for 24 hr in pancreatic islet media (PIMS, Prodo Laboratories). Single cells were dissociated with TrypLE for ~15-30 minutes, washed with PBS, stained with Calcein-AM/propidium iodide, and sorted onto nanoSPLITS chips using a cellenONE cell sorter. For the proteomic sample prep all steps utilized the nanoPOTS sample handling robot. Lysates were reduced with DTT, alkylated with IAA, digested overnight with trypsin/Lys-C at 37C, and vacuum dried / stored at -20C. LC-MS/MS data was acquired using TIFF DDA methodology on a Thermo Tribrid mass spectrometer. Data was searched using FragPipe version 20.0 before downstream analysis in R. [doi:10.25345/C5ZK55X61] [dataset license: CC0 1.0 Universal (CC0 1.0)]

Keywords: nanoSPLITS ; nanoPOTS ; single-cell ; multiomics ; multimodal ; scRNAseq ; scProteomics ; pancreas ; islets

Contact

Principal Investigators:
(in alphabetical order) 		James M. Fulcher, Pacific Northwest National Laboratory, USA
Submitting User: alchemistmatt

Publications

Fulcher JM, Markillie LM, Mitchell HD, Williams SM, Engbrecht KM, Degnan DJ, Bramer LM, Moore RJ, Chrisler WB, Cantlon-Bruce J, Bagnoli JW, Qian WJ, Seth A, Paša-Toli? L, Zhu Y.
Parallel measurement of transcriptomes and proteomes from same single cells using nanodroplet splitting.
Nat Commun. 2024 Dec 5;15(1):10614. Epub 2024 Dec 5.

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