A pixel arrangement structure, a display method and a preparing method of a pixel arrangement structure, and a display substrate are provided. The pixel arrangement structure includes first color sub-pixel blocks, second color sub-pixel blocks, and third color sub-pixel blocks. Each of the plurality of minimum repeating regions has a rectangular shape and includes a first virtual rectangle, and the first virtual rectangle includes one first color sub-pixel block, one second color sub-pixel block and one third color sub-pixel block. Any edge of the first virtual rectangle has a non-zero included angle with a first direction, and the first direction is a row direction or a column direction. The first color sub-pixel block is on a perpendicular bisector of the first edge, the second color sub-pixel block and the third color sub-pixel block are on different sides of the perpendicular bisector of the first edge.