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 to avoid cells stacking on top of each other. We suggest to first seed the adherent cells at a low density and allow them to adhere to the slide. Then, seed the suspension cells, also at a low density, and centrifuge prior to proceeding to the fixation step. During optimization, we strongly recommend taking images pre- and post-fixation, and leveraging one of our optimization kits: Teton Onboard Cell Paint Imaging Kit (860-00047), Teton Custom Antibody Screening Kit (860-00035) or Teton Cell Paint Probe Kit (830-00045).
When reviewing using a fluorescent microscope, ensure good borders around the cell edges and that the cells are not overlapping. Refer to the Teton Cell Culture Optimization Tech Note for more considerations, and the Cell Culture Hub for a repository of reference images.