Single Cell Track and Trace: live cell labelling and temporal transcriptomics via nanobiopsy

Author:

Marcuccio FabioORCID,Chau Chalmers C.ORCID,Tanner GeorgetteORCID,Elpidorou MarilenaORCID,Finetti Martina A.ORCID,Ajaib Shoaib,Taylor MoragORCID,Lascelles Carolina,Carr IanORCID,Macaulay IainORCID,Stead Lucy F.ORCID,Actis PaoloORCID

Abstract

ABSTRACTSingle-cell RNA sequencing has revolutionised our understanding of cellular heterogeneity, but whether using isolated cells or more recent spatial transcriptomics approaches, these methods require isolation and lysis of the cell under investigation. This provides a snapshot of the cell transcriptome from which dynamic trajectories, such as those that trigger cell state transitions, can only be inferred. Here, we present cellular nanobiopsy: a platform that enables simultaneous labelling and sampling from a single cell without killing it. The technique is based on scanning ion conductance microscopy (SICM) and uses a double-barrel nanopipette to inject a fluorescent dye and to extract femtolitre-volumes of cytosol. We used the nanobiopsy to longitudinally profile the transcriptome of single glioblastoma (GBM) brain tumour cells in vitro over 72hrs with and without standard treatment. Our results suggest that treatment either induces or selects for more transcriptionally stable cells. We envision the nanobiopsy will transform standard single-cell transcriptomics from a static analysis into a dynamic and temporal assay.

Publisher

Cold Spring Harbor Laboratory

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