Question: Can I use different permeabilization time points on a single Visium Gene Expression slide?
Answer: This depends upon whether H&E staining for morphological context or immunofluorescence staining for protein detection will be performed.
- H&E stained tissue sections
If the tissue sections are stained with H&E, different permeabilization time points can be performed on the same Visium Gene Expression Slide by following the method used during the Tissue Optimization workflow. The Permeabilization Enzyme should be added to the sections requiring the longest time first and work backward to the shortest incubation time. We recommend leaving the H&E stained sections dry until the permeabilization enzyme is added. We have learned that H&E staining has RNA stabilizing properties. Therefore, assay performance should not be affected when processing H&E stained sections on a single Gene Expression slide using different permeabilization time points.
- Immunofluorescence-stained tissue sections
For IF stained tissue sections, it is not recommended to process sections with more than one permeabilization time per Visium Gene Expression Slide. Immediately following IF imaging and Slide Cassette assembly, wash buffer should be added to the sections to prevent the tissue sections from drying out. Leaving IF stained tissue sections covered with wash buffer during the Gene Expression permeabilization step has not been tested and may impact assay performance.
Product: Visium for fresh-frozen