Laser-capture microdissection for spatial transcriptomics of immunohistochemically detected neurons.

ABSTRACT: We developed a versatile 'IHC/LCM-Seq' method for spatial transcriptomics of immunohistochemically detected neurons collected with laser-capture microdissection (LCM). IHC/LCM-Seq uses aluminon and polyvinyl sulfonic acid for inventive RNA-preserving strategies to maintain RNA integrity in free-floating sections of 4% formaldehyde-fixed brains. To validate IHC/LCM-Seq, we first immunostained and harvested striatal cholinergic interneurons with LCM. RNA preparations were subjected to random primer-based cDNA library preparation and bulk sequencing on the NextSeq Illumina platform. IHC/LCM-Seq detected ∼16,000 transcripts, reaching the sensitivity of a reference 'LCM-Seq method' developed for fluorescently tagged neurons microdissected from lightly formaldehyde-fixed and slide-mounted brain

SUBMITTER: Gocz B 

PROVIDER: S-EPMC11910328 | biostudies-literature | 2025 Feb

REPOSITORIES: biostudies-literature