Why does the foam spontaneously detach the day after seeding or when I add the medium?

There are many things that affect bad foam attachment or poor foam stability:

1. The surface for foam generation is not hydrophobic enough. Culture plates that are intended for suspension culture or are non-tissue treated (non-TC) are usually hydrophobic. We recommend using the plates from SARSTEDT, ref:83.3922.500.

2. The generated foam diameter is too small. If the area size of the foam is too small, that will result in a bigger lifting force once the medium is added. The foam size should be 0.7-1 cm in diameter for good attachment.

3. The silk solution has been standing at room temperature too long after thawing. The silk solution should be used directly after thawing (within 45-60 min). If the solution looks milky, do NOT spin/centrifuge the Biosilk solution as that will damage the product. 

4. The stabilization temperature has been lower than 37°C. Try only using the outer wells of the plate so that the stabilization temperature reaches 37°C faster. Alternatively, the stabilization time was not long enough (less than 20 min). Prolong the incubation time to 25 min.

5. After stabilization, carefully drop the medium both on the foam and around it, before slowly covering the foam with the medium. Filling up medium around the foam only will generate a lifting force to the foam. Do not add the medium too fast as that may break the foam apart.