The invention discloses a method of judging quality of a blastocyst by using a protein chip. The method comprises the following steps: I, when an embryo is developed to the blastocyst stage, transferring the blastocyst; II, dropping and absorbing a culture liquid to a sample tube and adding a protease inhibitor into the sample tube to prepare a to-be-tested sample; III, adding the to-be-tested sample into a 384 porous plate, adding a sample application liquid into each hole to prepare a to-be-sample applied sample; IV, dropping the to-be-sample applied sample in each hole to the chip, puttingthe chip in a wet box and drying the chip in an oven; V, incubating a protein antibody for the dried chip to prepare the protein chip; and VI, carrying out fluorescence detection on each protein chipand comparing the contents of six proteins of each blastocyst with standard contents of the six proteins. The method can evaluate the development condition of the blastocyst noninvasively, objectivelyand quantitatively, can be used for screening blastocysts which are high in quality and have planting potential and helps to improve the accuracy of screening the quality of the blastocyst.