The　formation　of　2D　colloid　nanosphere　templates　is　the　prerequisite　to　fabricate　nanoparticle　arrays　by　nanosphere　lithography.　A　new　fabrication　method　which　turns　colloidal　polystyrene　nanospheres　into　2D　crystals　directly　on　substrates　by　self-organization　was　presented.　Important　parameters　for　obtaining　large,　well-ordered　templates　are　discussed,　i.e.,　the　inclination　angle　of　substrates,　the　concentration　of　suspension,　the　physical　and　chemical　properties　of　substrate,　and　the　ambient　temperature　and　humidity.　The　samples　are　characterized　by　atomic　force　microscopy.　The　results　indicate　that　the　method　enables　the　fabrication　of　well-ordered　nanosphere　templates　with　large　area　up　to　square　centimeter,　which　is　suitable　for　the　consecutive　nanosphere　lithography　process.