Background: Deparaffinization is an important step in histological staining to remove paraffin before the staining process. Xylol is commonly used as a standard deparaffinizing agent; however, it has toxic effects on health and the environment. Virgin coconut oil is used as an alternative deparaffinizing agent because it is safer, environmentally friendly, and does not damage tissue. Research Objective: Determine the histological appearance of hematoxylin eosin staining in prostate cancer tissue deparaffinized using virgin coconut oil as an alternative agent compared with xylol as the standard. Research Methods: This study employed a cross-sectional approach. The samples consisted of archived prostate cancer tissue blocks divided into three treatment groups: deparaffinization using xylol, deparaffinization using virgin coconut oil at 50°C for 10 minutes, and deparaffinization using virgin coconut oil at 60°C for 8 minutes. The staining results were evaluated using a scoring method adopted from Ariyadi and Suryono (2017) based on the parameters of nuclear and cytoplasmic staining clarity, staining uniformity, and staining clearness of the preparations. The data were analyzed descriptively. Research Results: Microscopic observations showed that preparations deparaffinized using virgin coconut oil at 50°C for 10 minutes and 60°C for 8 minutes produced staining comparable to xylol. All preparations in the three groups achieved maximum scores, indicating clearly visible nuclei, evenly distributed and contrasting red cytoplasm, high staining clarity, and high color uniformity. Conclusion: Virgin coconut oil at 50°C for 10 minutes and 60°C for 8 minutes can be used as an alternative deparaffinization agent in hematoxylin eosin staining of prostate cancer tissue, showing good staining quality comparable to xylol. Keywords: Deparaffinization, Xylol, Virgin Coconut Oil, Hematoxylin Eosin Staining, Temperature and Time.