Multimodal single-cell profiling of the human heart maps cellular niches and reveals regulatory networks. A computational package also maps drug-target pairs to these single-cell data. NBThighlight
. Slices of 5–10 mm thickness from frozen tissue were first sectioned with a cryostat in a 50-μm thickness section. All sections from each sample were homogenized using a 7 ml glass Dounce tissue grinder set with 8–10 strokes of a loose pestle and 8–10 strokes of a tight pestle in homogenization buffer , 1× protease inhibitor, 0.4 U μlSUPERaseIn and 0.1% Triton X-100 in nuclease-free water). Homogenate was filtered through a 40 μm cell strainer .
For formalin-fixed paraffin-embedded samples, fresh samples were fixed in >5 times their volume of 4% v/v formalin at ambient temperature for 24 h before processing to paraffin on a Tissue-Tek Vacuum Infiltration Processor 5 . FFPE blocks were sectioned at 5 μm using a microtome . Samples of the microanatomical regions of interest were selected on the basis of morphology with expert review , orientation and either RNA integrity number or DV200 that was obtained using an Agilent 2100 Bioanalyzer. In addition, FFPE tissues were checked for possible detachment issues using 10x Genomics Adhesion test slides. FFPE Visium Spatial Gene Expression was performed following the manufacturer’s protocol.
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