Abstract: The advent of single-cell transcriptomics (scRNA-seq) has revolutionized biological and biomedical research by enabling the study of cellular diversity and states within a population. However, standard approaches often fail to capture lowly expressed genes and subtle signatures from rare cells, while also losing spatial and phenotypic information during library preparation. Chen et al. (2022) demonstrated a novel approach to single-cell transcriptome profiling by employing a biopsy technology that retrieves cytoplasmic material from live cells using FluidFM® (Fluidic Force Microscopy). Since these single-cell biopsies are not destructive to the cell, it becomes possible to detect the transcriptome while keeping the characterized cell alive, opening the possibility to temporal transcriptomics for the examination of long-term transcriptomic changes in single-cells. One such process is the differentiation of adipocyte stem and progenitor cells, which was successfully characterized with live-seq (Chen et al. (2022)). Here, we introduce a streamlined workflow using the FluidFM OMNIUM platform to collect cytoplasmic biopsies from Panc-1 cells and generate transcriptomes from less than a cell's worth of RNA. Utilizing the ultra-low RNA input LUTHOR™ HD library preparation kit, we show that transcriptomes can be prepared and sequenced from as little as 0.15 pg of RNA—equivalent to less than 1% of a Panc-1 cell. This advancement highlights the ability of the FluidFM OMNIUM platform, combined with LUTHOR HD, to enable high-resolution single-cell RNA profiling from live-cell biopsies, opening new avenues for investigating cellular heterogeneity and dynamics.
Chen, W., Guillaume-Gentil, O., Rainer, P.Y. et al. Live-seq enables temporal transcriptomic recording of single cells. Nature 608, 733–740 (2022). https://doi.org/10.1038/s41586-022-05046-9
