Protocol for preparing fresh frozen soybean tissues for spatial transcriptomics

Spatial transcriptomics is a disruptive technology that enables the identification of cell-type-specific transcripts en masse. However, the use of spatial transcriptomics for plant tissues has been challenging due to issues related to tissue preparation. Here, we present a protocol for preparing fresh frozen soybean tissues for spatial transcriptomics. We describe the steps for embedding, cryosectioning, fixation, staining, and imaging of soybean tissues. We then detail procedures for library preparation and sequencing. For complete details on the use and execution of this protocol, please refer to Zhang et al.¹.