An experiment was conducted in a greenhouse to determine the effects of different supplemental light spectra on the growth, nutrient uptake, and fruit quality of four strawberry cultivars. The plants were grown under natural light and treated with blue (460 nm), red (660 nm), and red/blue (3:1) lights. Results showed that the “Parous” and “Camarosa” had higher fresh and dry mass of leaves, roots, and crowns compared to the “Sabrina” and “Albion”. The use of artificial LED lights improved the vegetative growth of strawberry plants. All three supplemental light spectra significantly increased the early fruit yield of cultivars except for “Parous”. The red/blue supplemental light spectrum also increased the fruit mass and length of the “Albion”. Supplemental light increased the total chlorophyll in “Camarosa” and “Albion”, as well as the total soluble solids in fruits. The “Albion” had the highest concentration of fruit anthocyanin, while the “Sabrina” had the lowest. The use of supplemental light spectra significantly increased the fruit anthocyanin concentration in all cultivars. Without supplemental light, the “Camarosa” had the lowest concentration of K and Mg, which increased to the highest concentration with the use of supplemental light spectra. All three spectra increased Fe concentration to the highest value in the “Sabrina”, while only the red/blue light spectrum was effective on the “Camarosa”. In conclusion, the use of supplemental light can increase the yield and fruit quality of strawberries by elevating nutrients, chlorophyll, and anthocyanin concentrations in plants.