Adding the hydrogel as a dome shape is not recommended. The dome shape may not stick to the bottom of the culture plate for long term culture. The hydrogel should cover the whole bottom of the well plate. We suggest gently tilting/swirling the well plate after adding the hydrogel to ensure the gel coats the entire bottom of the well plate. Using a 96 well plate can reduce the usage volume of the hydrogel to 30-75 µL/well. There is no issue with molecular penetration when adding the cover medium to feed the cells.