When designing Teton™ experiments, there may be a situation where it makes sense to include both adherent and suspension cells on the same slide.
If the adherent and suspension cells are in separate wells, our recommendation is to grow the adherent cells on the slide until they reach the desired confluency. When you are ready to proceed, follow the Teton User Guide to Attach Suspension Cells in the empty wells. Once the suspension cells have been centrifuged and adhered, both can be fixed keeping in mind that the suspension fixation protocol requires adding 8% formaldehyde without removing the PBS resulting in 4% formaldehyde fixation. Adherent cells are fixed using 4% formaldehyde.
Co-culturing adherent and suspension cells will take more optimization, and we strongly recommend using a Teton Optimization Kit (860-00022) while optimizing your conditions. Ultimately, we want to avoid cells stacking on top of each other. To avoid this, we recommend seeding the adherent cells at a low density and, similarly, use the suspension cells at a low density to reduce the likelihood of suspension cells landing on the adherent cells. When reviewing using a fluorescent microscope, make sure you can see good borders around the cells and that the cells are not overlapping.