Raghubar, Arti M., Crawford, Joanna, Jones, Kahli, Lam, Pui Y., Andersen, Stacey B., Matigian, Nicholas A., Ng, Monica S.Y., Healy, Helen, Kassianos, Andrew J., and Mallett, Andrew J. (2023) Spatial Transcriptomics in Kidney Tissue. In: Hewitson, Tim D., Toussaint, Nigel D., and Smith, Edward R., (eds.) Kidney Research: Experimental protocols. Methods in Molecular Biology . Springer, New York, NY, USA, pp. 233-282.

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Abstract

Unlike bulk and single-cell/single-nuclei RNA sequencing methods, spatial transcriptome sequencing (ST-seq) resolves transcriptome expression within the spatial context of intact tissue. This is achieved by integrating histology with RNA sequencing. These methodologies are completed sequentially on the same tissue section placed on a glass slide with printed oligo-dT spots, termed ST-spots. Transcriptomes within the tissue section are captured by the underlying ST-spots and receive a spatial barcode in the process. The sequenced ST-spot transcriptomes are subsequently aligned with the hematoxylin and eosin (H&E) image, giving morphological context to the gene expression signatures within intact tissue. We have successfully employed ST-seq to characterize mouse and human kidney tissue. Here, we describe in detail the application of Visium Spatial Tissue Optimization (TO) and Visium Spatial Gene Expression (GEx) protocols for ST-seq in fresh frozen kidney tissue.

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