Purpose : Colonization of denture soft lining materials by Candida albicans can result in clinical problem, and deterioration of the materials. This study aimed to compare the retention and penetration of C. albicans into four denture soft lining materials commonly used. Materials and methods : Four denture soft lining materials (Coe-comfort$^{(R)}$, Coe-soft$^{(R)}$, GC soft liner$^{(R)}$, and Tissue conditioner$^{(R)}$) discs were prepared to glass slide and dental stone. Adherence of yeast to surfaces was monitored after one hour incubation of standardized washed cell suspension with test disc surfaces. Adherent cells stained with acridine orange were counted fluorescence microscopy. Penetration of yeast into materials bonded with acrylic resin after 1, 2, 3,4, 5, 6 and 7 days incubation was observed through sections stained using acridine orange and estimated to quantitative analysis using radioisotope. Results : There was statistical significance in cell numbers between smooth and rough surfaces(p<0.05). Higher numbers of cells were observed on rough surfaces. There was statistical significance in adherent cell numbers into smooth and rough surfaces individually(p<0.05). According to the increase of incubation periods, the cells penetrated into denture soft lining materials were shown to increase. The differences among all kinds of soft liner were statistically significant(p<0.05),and the largest number of cells penetrated into soft liners was observed in the Coe-soft$^{(R)}$. Conclusion : Initial adherence and penetration of yeast into denture soft lining materials has been influenced by surface roughness and chemical composition of them. The selection of appropriate materials and their fabrication may promote clinical performance.