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Conditioning of diluted exhaust gas by Thermo-Conditioner prior to measurement has been proposed by the GRPE/PMP Research Council of the United Nation in order to achieve stability in nano-particle measurement. In this study the effect of thermo-conditioner on the thermo-physical behavior of nano-particle under different conditions have been clarified. Stability in measurement was also attempted depending on the characteristics of nano-particles. Quality of the raw exhaust gas, the dilution ratio and temperature, and the thermal conditioning temperature were considered as the main parameters. Exhaust gas from a medium duty DI diesel engine was used for analysis. Scanning Mobility Particle Sizer was used for measuring the concentration of nano-particles. It was concluded that the concentration of nuclei-mode particles within the size range of 15∼30 nm are significantly influenced by the thermal conditioning temperature.