In this seminar, we will discuss Live-seq, a transformative method for single-cell gene expression profiling that eliminates the need for cell isolation and lysis. Live-seq utilizes Fluidic Force Microscopy (FluidFM) to extract cytosolic picolitre biopsies from living cells, which are subsequently analysed with a highly-sensitive RNA-seq protocol. While the resulting gene expression profiles faithfully represent lysed cell transcriptomes, the sampled cells remain alive and functional in their original microenvironment. Live-seq thus enables spatially-resolved, longitudinal profiling and phenotyping of individual cells, offering exciting opportunities for investigating cellular dynamics and cell-cell communication.