Journal: STAR Protocols

Article Title: Protocol to evaluate mouse brain spatial cell type-resolved transcriptomic discoveries using 10× Visium spatial transcriptomics and FLEX scRNA-seq

doi: 10.1016/j.xpro.2025.104277

Figure Lengend Snippet: Photographic illustration of steps involved in scRNAseq for tissue section fixation and dissociation (A) Tissue sections incubated in Fixation Buffer without curls being touched by the pipette. (B) After fixation, tissue sections either float to the top of the solution or pellet in the bottom of the tube. (C) Sections will pellet during centrifugation, remove supernatant and resuspend. (D) Addition of Dissociation Solution and mix/dissociate by pipette and transfer to Miltenyi C Tube. (E) Miltenyi C Tube flipped upside down and attached to Octo Dissociator and sample dissociated after gentleMACS program. (F) For adult samples and removal of debris clusters, filter dissociated tissue through Pre-Separation Filter (30 μm) placed on a 15-ml tube keeping the P1000 at an angle when pipetting through filter. (G) Successful dissociation of tissue sections stained with AOPI stain imaged on an EVOS microscope (10× magnification, scale bar = 300 μm) and counted with a hemocytometer.

Article Snippet: Add to respective Miltenyi C tubes.

Techniques: Incubation, Transferring, Centrifugation, Staining, Microscopy