The cross-impingement phenomenon always appears in several diesel engines with two or more injectors. Meanwhile, the application of biofuels has a great potential in realizing clean and efficient combustion. Therefore, the investigation aims to explore the cross-impingement characteristics at small (10%), middle (30%), and large (50%) biodiesel-butanol blended proportions. Experiments are conducted in a constant-volume combustion chamber with twin injectors. Spray images are captured by optical diagnosis techniques. Several macroscopic parameters are obtained, including diffusion length, collision width, and spray area. Results show that the cross-impingement accelerates the droplet interaction, and the spray presents a “fan-shaped” behavior after the collision, which promotes a more uniform mixing between the fuel and ambient gas. As the twin sprays collide at 120 deg, the vapor-phase vertical diffusion rate is close to the vertical component of the single spray, and the horizontal diffusion rate is about 1.2 times the vertical diffusion rate. The cross-impingement is likely to decrease the spray-wall impingement owing to a change in the diffusion direction. At various blended fuels, the biodiesel blended with 30% n-butanol displays the smallest liquid-phase diffusion length, width, and area. The further increase in the n-butanol mixing ratio leads to larger liquid-phase parameters. Contrary to the biodiesel blended with 10% n-butanol, the biodiesel blended with a higher proportion of n-butanol presents faster vapor-phase diffusion, which promotes fuel-gas mixing.