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This paper describes the sensitive cooling performance change of J-T refrigerator for cryosurgical probe due to its working fluid. The analytical results of using 50 bar nitrous oxide are compared with the case of 300 bar argon. Bio-heat equation is numerically solved to investigate the effect of the probe temperature and the cooling power of the J-T refrigerator. The refrigerator using 50 bar nitrous oxide has larger cooling power above 185 K than the one with 300 bar argon, which enables fast cooling at early stage of cryosurgery, but the biological tissue away from the probe tends to be cooled slowly after the probe reaches its lowest operating temperature. When the repeated freeze-thaw cycle is employed for main tissue destruction mechanism, using high pressure nitrous oxide is more advantageous than argon if the freezing operation is within 2-3 minutes. The probe with high pressure argon is more suitable for the case of longer freeze-thaw cycle with fewer repetitions.