The potential of human pluripotent stem cells as a renewable source of specialized cells for use in regenerative medicine has been limited by the lack of reproducible and robust differentiation protocols. The formation of embryoid bodies (EBs) is a critical intermediate in the induction of lineage-specific differentiation and is often necessary to derive mature, functional, specialized cell types. Current EB production methods form EBs inconsistently and produce EBs of varying sizes, resulting in ambiguous and often irreproducible data. A novel approach for reproducible production of size-controlled EBs has been developed. Brightfield imaging is combined with precise laser cutting of confluent human stem cell cultures into defined size cell sections to generate EBs of specific sizes. Laser-mediated EB generation was used to consistently produce uniform (7-14% CV), size-specific EBs from several human iPSC lines with high yield (>85%). Results show that EBs of specific sizes produced by laser-mediated EB generation differentiated more efficiently into multi-lineage specialized cell types; ~5-fold increase in the formation of neural progenitor cells and motor neurons, up to 15-fold increase in cardiomyocyte production, and >35-fold increase in the formation of hepatocyte-like cells when compared with EBs produced by typical methodology based on phenotypic and marker expression analysis. This approach provides an efficient, reproducible, and scalable method for EB generation which should enable development of universal, standardized procedures for the derivation of specialized cell types from human pluripotent stem cells.