Carver et al.,Aged Brain Spatial Profiling - GeoMx v1

This protocol provides the staining process for GeoMx digital spatial profiling of mouse brain used in in Carver et al., "Senescent- and disease-associated microglia are modifiable features of aged brain white matter". T Sections were cut at 5 μm from FFPE blocks. Brain slices were adhered to glass slides and stained directly on the surface The immunostaining process involves unconjugated- or fluorophore-conjugated- primary antibodies and secondary antibodies conjugated to fluorophores. Due to old brain accumulation of lipofuscin granules that autofluoresce, we note that in order to achieve maximal signal, a quenching step is performed to extinguish the autofluorescent noise. Transcriptome in situ hybridiazation was performed with the GeoMx Whole Transcriptome Atlas for mouse. Sections were imaged and processed on the GeoMx digital spatial profiling machine platform.