How can the meniscus be minimized in the ibidi µ-Plate 24 Well?

Meniscus formation is an intrinsic property of small open wells and cannot be completely avoided.

A meniscus is more likely to build up on a hydrophilic surface than on a hydrophobic surface. Since most adherent cell types do not attach well onto hydrophobic surfaces, ibidi offers culture vessels with a hydrophilic, tissue culture-treated surface and optimal cell adhesion properties (ibidi µ-Plate 24 Well). However, in this case, meniscus formation can be more prominent than when using an uncoated plate.

To reduce meniscus formation, try using the ibidi µ-Plate 24 Well Uncoated, which has an untreated, hydrophobic surface. When filled with water, this surface allows almost no meniscus to build up. However, the proteins of the cell culture medium will attach to the surface of the well. After some time, this will lead to a non-covalent, monomolecular coating that will create a hydrophilic surface and a prominent meniscus. Neither water nor buffer can wash off the protein coating.

To minimize meniscus issues during imaging AND make sure that the cells attach, you can try the following protocol in the ibidi µ-Plate 24 Well Uncoated:

1. Coat the wells with binding protein (e.g., Poly-L-lysine or fibronectin) to support cell attachment. Use the lowest volume possible to ensure that only the lower part of the well is hydrophilic (e.g., 0.5–0.8 ml). Handle the plate with care so that the upper part of the well is not in contact with the protein solution. After coating, carefully remove the solution.
2. Seed the cells in 0.5–0.8 ml of culture medium. Check regularly, at least once a day, to see if this amount is sufficient to ensure the survival of your cells. Replace the medium, if necessary, without getting the upper part of the well 'wet'.
3. For imaging, replace the culture medium with 1.5–2 ml of protein-free solution or buffer. Now, the hydrophilic coating from the first step should be below your filling level, and only a minimal meniscus should have formed due to the hydrophobicity of the upper parts of the well.

Please note: We have not tested this protocol ourselves, and therefore recommend that you try this approach with a free sample.

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