Microalgae cells have been recognized as a promising sustainable resource to meet worldwide growing demands for renewable energy, food, livestock feed, water, cosmetics, pharmaceuticals, and materials. In order to ensure high-efficiency and high-quality production of biomass, biofuel, or bio-based products, purification procedures prior to the storage and cultivation of the microalgae from contaminated bacteria are of great importance. The present work proposed and developed a simple, sheathless, and efficient method to separate microalgae Chlorella from bacteria Bacillus Subtilis in a straight channel using the viscoelasticity of the medium. Microalgae and bacteria migrate to different lateral positions closer to the channel centre and channel walls respectively. Fluorescent microparticles with 1 μm and 5 μm diameters were first used to mimic the behaviours of bacteria and microalgae to optimize the separating conditions. Subsequently, size-based separation in Newtonian fluid and in viscoelastic fluid in straight channels with different aspect ratios was compared and demonstrated. Under the optimal condition, the removal ratio for 1 μm microparticles and separation efficiency for 5 μm particles can reach up to 98.28% and 93.85% respectively. For bacteria and microalgae cells separation, the removal ratio for bacteria and separation efficiency for microalgae cells is 92.69% and 100% respectively. This work demonstrated the continuous and sheathless separation of microalgae from bacteria for the first time by viscoelastic microfluidics. This technique can also be applied as an efficient and user-friendly method to separate mammalian cells or other kinds of cells.