The invention discloses a quantitative high-vitality yeast cell screening method based on a TTC staining method. The quantitative high-vitality yeast cell screening method comprises the steps as follows: coating a plate with a yeast cell liquid to be detected, and culturing the yeast cell liquid until single colonies grow; then inoculating the yeast cell liquid in a culture solution in each hole of a 96-hole cell culture plate, culturing the yeast cell liquid until OD values of thalli obtained through measurement are in a range of 1-3, centrifuging the liquid and abandoning supernatant; adding a TTF extracting agent to each hole of the 96-hole cell culture plate, and oscillating the liquid for 10-15S before centrifugation; and absorbing oscillated and centrifuged supernatant in a 96-hole EKISA plate, measuring a TTF absorption value under the wavelength of 486nm through an EKLSA reader, and calculating the content of TTF in each hole according to a TTF standard curve and the measured cell OD values to determine the vitality of cells. Screening results reflect the vitality of the cells directly and reliably, the screening time can be reduced obviously, and human errors caused by visual judgments during an operation process can also be reduced.