Journal: bioRxiv
Article Title: MERFISH 2.0, an ultra-sensitive single-cell spatial transcriptomics imaging chemistry across diverse tissue types
doi: 10.64898/2026.03.06.710199
Figure Lengend Snippet: Formalin-fixed paraffin-embedded (FFPE) human tissues, including breast cancer specimens and lung and breast cancer tissue microarrays (TMAs), were profiled using the 815-plex Human Breast Cancer Panel or the Human Immuno-Oncology Panel following the MERFISH 1.0 and MERFISH 2.0 workflows. A) Sensitivity comparison between MERFISH 1.0 and 2.0 across FFPE human samples. RNA transcript counts per 100 µm² were used as a metric of sensitivity. Each condition includes at least 3 biological replicates (N=3). MERFISH 2.0 consistently increases the sensitivity for transcripts detection across samples. B) Concordance analysis between MERFISH 1.0 and MERFISH 2.0 measurements within each tissue. Person correlation coefficients calculated across samples demonstrate strong agreement between the two chemistries (r > 0.8). C) . UMAP visualization of single-cell transcriptomic profiles from FFPE human breast cancer displaying the different major cell types identified with MERFISH 1.0 (left), and MERFISH 2.0 (right). A total of 367,662 cells passed QC for MERFISH 1.0 and 654,332 cells passed QC for MERFISH 2.0. D) Transcript detection across individual TMA cores. Top, histograms showing the distribution of transcripts per cell for MERFISH 1.0 and MERFISH 2.0 across cores (R1–R5), with mean transcript counts indicated. Bottom, spatial distribution of Leiden clusters across tissue cores detected with MERFISH 1.0 (middle) and 2.0 chemistry (bottom), demonstrating much greater transcript coverage and cell recovery with MERFISH 2.0. E) Concordance between MERFISH-derived cell type profiles and reference single cell RNA sequencing data. Heatmap and summary correlation analysis show improved agreement with scRNA-seq when using MERFISH 2.0 compared to MERFISH 1.0. F) Violin plot showing the dynamic range of RNA transcripts counts per cell across representative cell types identified in human breast cancer.
Article Snippet: MERFISH 1.0 sample preparation followed published protocols and manufacturer’s instructions: MERSCOPE Fresh and Fixed Frozen Tissue Sample Preparation User Guide (Vizgen PN 91600002) and MERSCOPE FFPE Tissue Sample Preparation User Guide (Vizgen PN 91600112).
Techniques: Formalin-fixed Paraffin-Embedded, Comparison, Single Cell, Cell Recovery, Derivative Assay, RNA Sequencing