Abstract

Single-cell RNA sequencing has revolutionised our understanding of cellular heterogeneity. However, conventional methods involve cell lysis and lack the ability to directly investigate dynamic trajectories underlying cellular state transitions, often relying on inference. Recently, Live-seq pioneered the temporal mRNA analysis of the same cell over two days via longitudinal cytoplasmic extraction using fluidic force microscopy. Here, we present a nanobiopsy platform that enables simultaneous injections and cytoplasmic sampling from an individual cell and its progeny without killing it. The technique is based on scanning ion conductance microscopy (SICM) and employs a double-barrel nanopipette to introduce exogenous molecules and longitudinally profile the transcriptome of individual glioblastoma (GBM) brain tumour cells in vitro over 72hrs, with and without standard treatment, including chemotherapy and radiotherapy. Our results suggest that treatment either induces or selects for more transcriptionally stable cells. We envision the nanobiopsy will contribute to the new emerging field that will transform standard single-cell transcriptomics from a static analysis into a dynamic and temporal assay. Top of Form