Particle　patterning　and　hopping　has　attracted　much　attention　owing　to　their　extensive　involvement　in　many　physical　and　biological　studies.　Here,　by　configuring　an　intriguing　Optofluidic,　we　are　able　to　pattern　500　nm　particles　into　a　2D　array　in　the　flow　stream.　We　also　achieve　a　2D　patterning　of　cryptosporidium　in　the　microchannel.　By　investing　particle-particle　interactions,　we　studies　the　long　ignored　new　particle　hopping　mechanisms,　and　used　them　to　screen　antibodies.　Our　observed　particle　hopping　in　the　flow　stream　completes　the　family　of　particle　kinetics　in　optofluidic　potential　wells　and　inspires　new　minds　in　the　develop　new　light　fields　in　the　microchannel.　The　2D　patterning　of　particles　facilites　the　parallel　culture　and　study　of　multiple　biological　samples　in　the　flow　stream.　©　2018　SPIE.