The research aims to determine the best type of medium and 2,4-D concentration on somatic embryo induction of Vanda tricolor tissue culture. The research was conducted in December 2018 to March 2019 at the Tissue Culture Laboratory of Agriculture Faculty, Universitas Muhammadiyah Yogyakarta. The research was conducted with a single factor experimental method and was arranged in a Completely Randomized Design (CRD) with 3 replications. The treatment tested was a combination of medium types (NDM and POC) with 2,4-D concentrations (0; 1; 3; 5 mg/l). TDZ 0,5 mg/l was added to each treatment. Parameters observed included the percentage of live explants, percentage of browning explants, percentage of contaminated explants, percentage of vitrified explants, time appears of pro-embryo, total of pro-embryos, percentage of callus explants, time appears of callus, callus diameter, callus texture, explant height, total of leaves, leaf color, and microscope observation. The results showed that the use of POC medium with a concentration of 1 mg/l 2,4-D tended to give the best effect as indicated by the time appears of pro-embryo at 2,72 weeks; total of pro-embryos as much as 2,44 pro-embryos; the percentage of callus explants of 0,84%; time appears of callus at 30,50 days; and callus diameter of 3,13 mm.