Photo-isolation chemistry for high-resolution and deep spatial transcriptome with mouse tissue sections
Mizuki Honda, Ryuichi Kimura, Akihito Harada, Kazumitsu Maehara, Kaori Tanaka, Yasuyuki Ohkawa, Shinya Oki
Abstract
Photo-isolation chemistry (PIC) enables isolation of transcriptome information from locally defined areas by photo-irradiation. Here, we present an optimized PIC protocol for formalin-fixed frozen and paraffin mouse sections and fresh-frozen mouse sections. We describe tissue section preparation and permeabilization, followed by in situ reverse transcription using photo-caged primers. We then detail immunostaining and UV-mediated uncaging to the target areas, followed by linear amplification of uncaged cDNAs, library preparation, and quantification. This protocol can be applied to various animal tissue types. For complete details on the use and execution of this protocol, please refer to Honda et al. (2021).