Disclosed is an electrofluidic support plate and a method for preparing the same, and an electrofluidic device comprising the support plate. The method comprises the following steps of: providing a substrate which has a surface provided with an electrode layer; arranging a first amorphous fluoropolymer layer on the surface of the substrate, and carrying out hydrophilic modification on a surface of the amorphous fluoropolymer layer; arranging pixel walls on the amorphous fluoropolymer layer after hydrophilic modification; arranging a second amorphous fluoropolymer layer which is a hydrophobic layer; the second amorphous fluoropolymer layer covering all surfaces of the pixel walls and a groove area encircled by the pixel walls; filling the groove area encircled by the pixel walls with a protective material; removing the second amorphous fluoropolymer layer not covered by the protective material and on a top of the pixel walls; and removing the protective material.