Objectives: This research was conducted to look for the vest conditions of Hematoxylin-eosin staining. Methods/ Statistical Analysis: Method was investigated the Hematoxylin-eosin staining, Microscopy and RGB analysis. Findings: The result was as follows. The thickness of the optimal condition is 3 cm, the time of the optimal Hematoxylin 4 minutes 30 seconds and eosin 3 minutes 45 seconds. Also, RGB analysis was green. So green showed a significant difference in the change width to each stage, and the biggest change was 52.74 at 2㎛ and 3㎛. Improvements: Therefore, an appropriate fixed time is needed to be found to get staining intensity and sharpness, and the freshness of reagent for each stage should be maintainedKeywords: Eosin, Hematoxylin, RGB, Microscopy, Stain