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Scalable in situ single-cell profiling by electrophoretic capture of mRNA

By Lars Borm, Alejandro Mossi Albiach, Camiel CA Mannens, Jokubas Janusauskas, Ceren Özgün, David Fernández García, Rebecca D Hodge, Ed S. Lein, Simone Codeluppi, Sten Linnarsson

Posted 13 Jan 2022
bioRxiv DOI: 10.1101/2022.01.12.476082

Methods to spatially profile the transcriptome are dominated by a trade-off between resolution and throughput. Here, we developed a method named EEL FISH that can rapidly process large tissue samples without compromising spatial resolution. By electrophoretically transferring RNA from a tissue section onto a capture surface, EEL speeds up data acquisition by reducing the amount of imaging needed, while ensuring that RNA molecules move straight down towards the surface, preserving single-cell resolution. We applied EEL on eight entire sagittal sections of the mouse brain and measured the expression patterns of up to 440 genes to reveal complex tissue organisation. Moreover, EEL enabled the study of challenging human samples by removing autofluorescent lipofuscin, so that we could study the spatial transcriptome of the human visual cortex. We provide full hardware specification, all protocols and complete software for instrument control, image processing, data analysis and visualization.

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