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

    Research output: Contribution to journalArticlepeer-review

    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).

    Original languageEnglish
    Article number101346
    JournalSTAR Protocols
    Volume3
    Issue number2
    DOIs
    Publication statusPublished - Jun 17 2022

    All Science Journal Classification (ASJC) codes

    • General Immunology and Microbiology
    • General Biochemistry,Genetics and Molecular Biology
    • General Neuroscience

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