Many factors contribute to the reluctance towards blood donation, but available studies done in Malaysia involving University students does not reflect the knowledge of the public in general. The objective of this study is to determine knowledge, attitude practice towards blood donation among the Sandakan population. A cross-sectional study was employed using an adapted 29-item structured validated questionnaire available in English and Bahasa, consisting of subject’s demography, questions regarding knowledge, attitude, and perception of blood donation. Convenient random sampling was done within the hospital compound, 79 healthy adults consented, and their data were used for the final data analysis, yielding an excellent internal consistency (Cronbach’s α coefficient = 0.816). Out of all, 74.7% of the respondents had a high level of knowledge, and independent t-tests showed that those who were not married, had tertiary education, donated blood in the past, had a statistically significant higher level of knowledge and 96.2% of respondents have a positive attitude. Some donors (40.6%) donated blood for moral satisfaction, and only a quarter (25%) experienced adverse events. Fear of pain, needle, fainting was the highest reason for reluctance in blood donation (36.2% of non-donors), followed by self-perception of being medically unfit to donate (31.9% of non-donors). Even though the sampled population in Sandakan showed an adequate level of knowledge as well as a positive attitude towards blood donation, blood product shortage is still present. This study may contribute by serving as an educational platform for awareness and education to improve the number of blood donors.