%0 Journal Article
%T Open-source, high-throughput targeted in situ transcriptomics for developmental and tissue biology.
%A Lee H
%A Langseth CM
%A Salas SM
%A Sariyar S
%A Metousis A
%A Rueda-Alaña E
%A Bekiari C
%A Lundberg E
%A Garcı A-Moreno F
%A Grillo M
%A Nilsson M
%J Development
%V 151
%N 16
%D 2024 Aug 15
%M 39099456
%F 6.862
%R 10.1242/dev.202448
%X Multiplexed spatial profiling of mRNAs has recently gained traction as a tool to explore the cellular diversity and the architecture of tissues. We propose a sensitive, open-source, simple and flexible method for the generation of in situ expression maps of hundreds of genes. We use direct ligation of padlock probes on mRNAs, coupled with rolling circle amplification and hybridization-based in situ combinatorial barcoding, to achieve high detection efficiency, high-throughput and large multiplexing. We validate the method across a number of species and show its use in combination with orthogonal methods such as antibody staining, highlighting its potential value for developmental and tissue biology studies. Finally, we provide an end-to-end computational workflow that covers the steps of probe design, image processing, data extraction, cell segmentation, clustering and annotation of cell types. By enabling easier access to high-throughput spatially resolved transcriptomics, we hope to encourage a diversity of applications and the exploration of a wide range of biological questions.