In the process of pL micro-volume adhesive distribution,the itinerary of the transfer needle is repetitive,and the work is inefficient.To solve this problem,machine vision and improved GA-ACO algorithm were used to plan the path quickly and accurately,and improve the efficiency.The experimental platform of micro-volume adhesive distribution was built which consisted machine vision system,adhesive distribution system and drive control system.The location information of the fixed point on the acquired image was gotten by template matching.The path planning of micro-volume adhesive distribution was completed.Finally,the performance of the designed algorithm was compared with common algorithms.The results show that based on machine vision and improved GA-ACO algorithm,the volume of transfer adhesive reaches the pL level;the average diameter of the adhesive is 68.6 μm,and the adhesive falls exactly at the preset point;the itinerary of the transfer needle is cut by 67.9%,the working time is reduced drastically.It can improve the efficiency of pL micro-volume adhesive distribution significantly.