The invention discloses a multi-factor integrated screening plate, a manufacturing tool and a manufacturing method thereof. The manufacturing tool of the multi-factor integrated screening plate is composed of one or more spacer rings; the height of spacer rings is above 5mm; the spacer rings are used in a culture dish; the interior of the culture dish is divided into a plurality of zones by the spacer rings; the thickness of spacer rings is 1-5mm; the interior of the culture dish is divided into zones at equal distances or of equal area by the spacer rings; the spacer rings are made from stainless steel, aluminum alloy, polypropylene, polyethylene, polystyrene, polyvinyl chloride or glass; the spacer rings are prepared in the manner of mold processing or 3D printing. The multi-factor integrated screening plate is prepared by using the manufacturing tool. The multi-factor integrated screening plate is cut into areas under different screening conditions by the manufacturing tool.