Objective: To develop a scale in order to investigate the knowledge and attitudes toward homosexuality among people particularly at least high school graduated who live in Turkey and/or speak Turkish; and to conduct reliability and validity studies of this scale. Design and Methods: Homosexuality Attitudes Scale is a 62-item Likert-type scale (5 points) with response options labeled strongly disagree, disagree, neither agree nor disagree, agree, strongly agree. The items on the scale were selected from initial pool of 103 items written by experienced psychiatrists-sex therapist or derived from literature review and interviews with experts knowledgeable about homosexuality. After reviewing for readability by eight high school or university graduated people, we agreed 62 of the original items for inclusion in the scale. The scale was administered to 127 hospital workers and 109 of them completed the scale. Additionally, 37 of the sample completed Turkish version of Hudson and Ricketts''Attitudes toward Homosexuals Scale'. An item analysis was conducted to identify the statements that could best discriminate high and low scorers. Findings/Results: Reliability: In a sample of 109 hospital workers the 62-item scale demonstrated excellent internal consistency (Cronbach's alpha = 0.952). Six items had lower internal consistency, When these items were excluded, Cronbach's alpha for the 56-item scale was 0.954. Validity: In the sample described above, scores on our scale were positively correlated with Turkish version of Hudson and Ricketts' scale "Attitudes toward Homosexuals" scale (r = 0.882, p < 0.001). Conclusions: The Homosexuality Attitudes Scale is a reliable and valid scale in assessing knowledge and attitudes toward homosexuality and it can be used in related research.