Background: Scutellaria baicalensis is one of the most popular medicinal plants, which roots extracts are widespread in medicine and cosmetology. The area of growth of the S. baicalensis is rapidly declining; therefore, the involvement of biotechnological approaches to obtain its biomass is relevant. Since the content of flavones in cultured in vitro cells is usually much lower than in intact plants, there is a need to strengthen the synthesis of target substances. Objectives: The objective of this study is to investigate the effect of blue, white, and red light for the growth and content of the four main flavones (baicalin, wogonoside, baicalein, and wogonin) in the callus tissue of the S. baicalensis. Materials and Methods: Calluses in the experiments were continuously illuminated for a month with blue (420–480 nm), red (600–650 nm), warm white (400–800 nm) using LEDs with illumination of 1 μmol/m2/s. High-performance liquid chromatography was used to determine the content of the main flavones. Results: The presence of the light of all studied parts of the spectrum contributed to the elongation of the stationary phase against the background of callus growth suppression, in addition, the content of flavones increased, mainly due to baicalin. The maximum number of flavones was formed in blue light (5%). Conclusion: The blue light is an important factor to the accumulation of the main flavones in the calluses of S. baicalensis. The results obtained can be used not only in fundamental research but also in practice.