Abstract: Abstract: Breathing is an important physiological activity of egg as a living organism, and it has a great influence on its storage, conveyance and incubation. Therefore, it is of great significance to study egg's breathing regularity and respiration intensity in a day and during its storage time in a month. In this paper, non-invasive micro-test technique (NMT) was used to real-time monitor O2 flux passing in and out the eggs through the eggshell and study the breathing regularity and the respiration intensity of the eggs. With this method, gas exchange (O2) was measured and it was confirmed that breathing indeed exists. In order to study egg's respiration in a day, a customized micro-testing examination platform was constructed for the real-time monitoring of the intensity and rate of oxygen exchange between an egg and its external environment at 4 time points (6:00, 12:00, 18:00, and 24:00). On the other hand, egg's O2 flux was measured at the same time in a day during a month as to seeking for breathing regularity in its storage time. It was indicated that the optimal distance of probe towards the egg surface was 50 μm and the blunt end of the egg was selected as the optimum measurement location via the pre-experiment. The results show that there are three breathing patterns of eggs, consuming O2, exhaust O2, and both including consuming and exhausting O2. After group A testing, it is concluded that both fertilized and non-fertilized eggs breathe strong at dawn and weak at noon regularly in a day and the respiration intensity of the two types of eggs perform difference at different time points. Fertilized eggs have more active physical activity and the process of metabolism than those of non-fertilized ones. Fertilized egg respiration shows significant difference (P＜0.05) at noon and dawn in a day, but non-fertilized egg does not. In addition, after group B testing, it is intended that during storage time in a month, there exists two breathing peaks in its storage period in a month, and each peak appears at the eggs quality changing time. The second peak value exceeds the first peak value. It reveals that there's a unique breathing pattern of eggs that O2 respiration rate is high when life metabolism is active, and low when life metabolism is negative. Eggs laid lately are breathing relatively positive. This study can provide a reference for optimizing storage conditions, improving the hatchability of fertilized eggs and hatching egg embryo in further research.