Display substrate, display device, high-precision metal mask are provided. Display substrate includes: first, second, and third sub-pixels; in first direction, first and third sub-pixels are alternately arranged to form first sub-pixel rows, second sub-pixels form second sub-pixel rows; in second direction, first and second sub-pixel rows are alternately arranged; two first and two third sub-pixels in two adjacent rows and two adjacent columns form 2*2 array; in the array, two first sub-pixels are in different rows and in different columns, so are the two third sub-pixels, connection lines of centers of two first and two third sub-pixels form virtual quadrilateral, second sub-pixel is within virtual quadrilateral; for multiple distances from centers of two first and two third sub-pixels corresponding to same virtual quadrilateral to center of second sub-pixel, at least two distances are different. Brightness centers of virtual pixels have more uniform distribution.