1. Scalable in situ single-cell profiling by electrophoretic capture of mRNA using EEL FISH.
- Author
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Borm LE, Mossi Albiach A, Mannens CCA, Janusauskas J, Özgün C, Fernández-García D, Hodge R, Castillo F, Hedin CRH, Villablanca EJ, Uhlén P, Lein ES, Codeluppi S, and Linnarsson S
- Subjects
- Humans, Animals, Mice, RNA, Messenger genetics, In Situ Hybridization, Fluorescence methods, Eels genetics, Gene Expression Profiling methods, RNA analysis, Transcriptome genetics
- Abstract
Methods to spatially profile the transcriptome are dominated by a trade-off between resolution and throughput. Here we develop a method named Enhanced ELectric Fluorescence in situ Hybridization (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 toward the surface, preserving single-cell resolution. We apply EEL on eight entire sagittal sections of the mouse brain and measure the expression patterns of up to 440 genes to reveal complex tissue organization. Moreover, EEL can be used to study challenging human samples by removing autofluorescent lipofuscin, enabling the spatial transcriptome of the human visual cortex to be visualized. We provide full hardware specifications, all protocols and complete software for instrument control, image processing, data analysis and visualization., (© 2022. The Author(s).)
- Published
- 2023
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