Abstract The purpose of the study was to investigate the playfulness and the humor sense on a comparative analysis of different personalities. The samples of this study were by employment service in thirty organizations on questionnaire. The measure instruments used in this study include ¡§Adult Playfulness Scale¡¨, ¡§Multidimensional Sense of Humor Scale¡¨, ¡§Locus of Control Scales¡¨ and ¡§PANAS Scales¡¨. The samples of this study were 1213 employees. The results of survey were analyzed by one-way ANOVA, descriptive statistics analysis, reliability analysis, and factor analysis. The results of the study were as follows: 1. There were significant differences in the playfulness of the employees under the different sex, age, marriage status, job position, Type A personality, Locus of Control personality, Positive Affectivity, and Negative Affectivity. 2. There were significant differences in the humor sense of the employees under the different sex, age, marriage status, Locus of Control Personality, Positive Affectivity, and Negative Affectivity. 3. There were comparative analysis the highest trend between the playfulness and the humor sense on employees of sex for male, age under thirty, graduated above mater degree, Control Personality, the high tread Positive Affectivity, and the low tread Negative Affectivity.