GSM8908344: DKD 16; Homo sapiens; RNA-Seq
Routine FFPE sections of kidney tissue are deparaffinised, hydrated and antigenically repaired before overnight incubation with a pre-designed RNA hybridisation probe (WTA) that, in addition to containing a portion that binds to the target RNA, also joins a section of DNA sequence with a specific label via a UV-dissolvable linker moiety. The next day, a polychromatic fluorescent antibody (Nephrin, AQP1) was incubated to enable morphological observation of the tissue and to guide the selection of subsequent ROIs. Subsequent UV irradiation of selected ROIs results in the release of DNA oligonucleotide strands with photolysable linkers, which are then absorbed by capillary tubes into 96-well plates. The collected DNA oligonucleotide sequence is composed of a section of RNA target-specific marker, specific molecular tag UMI and primer binding site for RNA identification, spatial position reduction and splitting of the sample after sequencing.