Project description:Single-cell RNA sequencing (scRNA-seq) has revolutionized our understanding of cellular diversity, but remains constrained by scalability, high costs, and the destruction of cells during analysis. To overcome these challenges, we developed STAMP (Single-Cell Transcriptomics Analysis and Multimodal Profiling), a highly scalable approach for the profiling of single cells. By leveraging transcriptomics and proteomics imaging platforms, STAMP eliminates sequencing costs, enabling single-cell genomics of hundreds to millions of cells at an unprecedented affordability. Immobilizing (‘stamping’) cells in suspension onto imaging slides, STAMP supports single-modal (RNA or protein) and multimodal (RNA, protein and H&E) profiling, while retaining cellular structure and morphology. Its flexible, ultra-high-throughput formats facilitate the analysis of single or multiple samples in the same experiment, enhancing experimental scalability and adaptability. We demonstrate STAMP's versatility across diverse experimental contexts, including the profiling of peripheral blood mononuclear cells (PBMCs), cell lines and stem cells. We also stamped cells and nuclei from dissociated tissues from mouse organs to simulate the generation of cell atlases. Accessibility was further enlarged by analyzing nuclei from archival formalin fixed and paraffin embedded (FFPE) tissue samples. Combining RNA and protein profiling, we applied STAMP for high-throughput immuno-phenotyping of millions of blood cells, providing multimodal insights into cellular heterogeneity. We highlight the capability of STAMP to identify ultra-rare cell populations, simulating clinical applications for detecting circulating tumor cells (CTCs). By capturing lineage dynamics during stem cell differentiation and subtle changes in in vitro activated PBMCs, we further showed its utility for large-scale perturbation studies. These results validate STAMP as a first-of-its-kind single-cell imaging analysis strategy. We present data for 10,962,092 high quality cells/nuclei and 6,030,429,954 high quality transcripts. By replacing sequencing with imaging, STAMP enables high-resolution cellular profiling that is more accessible, scalable, and cost-effective. STAMP has the potential to transform our ability to map biological diversity and dynamics, significantly advancing research and clinical applications.

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Multimodal measurement of the transcriptome and proteome in G2/M arrested single cells using nanoSPLITS

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Parallel measurement of transcriptomes and proteomes from same single cells using nanodroplet splitting.

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