作者
Naomi Martin,Paul Olsen,Justin Quon,Jazmin Campos,Nasmil Valera Cuevas,J. S. Nagra,Marshall VanNess,Zoe Maltzer,Emily Gelfand,A. Oyama,Amanda Gary,Yimin Wang,Angela Alaya,Augustin Ruiz,Cade Reynoldson,Cameron Bielstein,Alice Pom,Cindy Huang,Cliff Slaughterbeck,Elizabeth Liang,Jason Alexander,Jeanelle Ariza,Jocelin Malone,J.L Melchor,Kaity Colbert,Krissy Brouner,Lyudmila Shulga,Melissa Reding,P.J. Latimer,Raymond E. A. Sánchez,Stuard Barta,Tom Egdorf,Zachary Madigan,Chelsea M. Pagan,Jennie Close,Brian Long,Michael Kunst,Ed S. Lein,Hongkui Zeng,Delissa McMillen,Jack Waters
摘要
Image-based spatial transcriptomics platforms are powerful tools often used to identify cell populations and describe gene expression in intact tissue. Spatial experiments return large, high-dimension datasets and several open-source software packages are available to facilitate analysis and visualization. Spatial results are typically imperfect. For example, local variations in transcript detection probability are common. Software tools to characterize imperfections and their impact on downstream analyses are lacking so the data quality is assessed manually, a laborious and often a subjective process. Here we describe imperfections in a dataset of 641 fresh-frozen adult mouse brain sections collected using the Vizgen MERSCOPE. Common imperfections included the local loss of tissue from the section, tissue outside the imaging volume due to detachment from the coverslip, transcripts missing due to dropped images, varying detection probability through space, and differences in transcript detection probability between experiments. We describe the incidence of each imperfection and the likely impact on the accuracy of cell type labels. We develop MerQuaCo, open-source code that detects and quantifies imperfections without user input, facilitating the selection of sections for further analysis with existing packages. Together, our results and MerQuaCo facilitate rigorous, objective assessment of the quality of spatial transcriptomics results.