Question: What should I do if tissue remains on the Visium Tissue Optimization slide after the tissue removal step?
Answer: You should not expect to see incomplete tissue removal for the tissues we have on the Optimized Tissue List when following the recommendations in our user guide:
- OCT brand, Sakura Tissuetek
- Methanol fixation
- Tissue Removal Buffer inspected to ensure no signs of precipitation before use
- Bubbles not introduced during dispensing Tissue Removal Mix into wells
- 2x SSC + 1% SDS pre-warmed to 50C prior to performing washes
Occasionally some tissue might remain in the no permeabilization control where permeabilization enzyme has not been added. However, in-house we have not observed autofluorescence from the remaining tissue. Therefore, if residual tissue only remains the no permeabilization control well, we recommend proceeding to fluorescent imaging.
If other time points display tissue remaining, we would recommend repeating the 2x SSC + 1% SDS washes ensuring the buffer is pre-heated to 50C prior. Then followed by the .2x and .1x SSC washes and final drying step. Then proceed to imaging. Rewashing the slide often helps if little tissue remains on the slide.