Studies have reported that homosexual patients fear they will not receive adequate care if they openly show their sexual orientation, for example, when introducing their partner. The aims of this study were to investigate the emotions of nursing staff and nursing students, and possible relations to cultural background and gender, towards homosexual patients; whether nursing staff and nursing students would choose to refrain from nursing homosexual patients, if the option existed; and, if so, how they express their wish to refrain from nursing this group of patients. All participants received verbal and written information before the study started. Returning a completed questionnaire indicated a participant's tacit consent. Approval was obtained from the heads of departments and persons in charge of nursing and nursing assistant programmes. The study had a descriptive, comparative design, and an Affect Adjective Checklist (AAC) and specially designed Nursing Behaviour Questionnaire (NBQ) were used. The participants included nurses and assistant nurses from an infectious disease clinic, and students enrolled in a university nursing programme and upper secondary assistant nurses' training, all in central Sweden. The findings showed that both professional nursing staff (response rate 67%, n = 57), and students (response rate 62%, n = 165), expressed emotions of homophobic anger, homophobic guilt and delight. Groups with a cultural background other than Swedish expressed more homophobia. No gender differences were indicated for homophobic emotions. In the professional group, 36% would refrain from nursing for homosexual patients if given the option. The corresponding figure for the students was 9%. The limitations were that the sample was small and not randomly selected, and as participation was anonymous no follow-up could be done. It was concluded that the emotional factors of homosexual anger and homosexual guilt might be of value in helping to explain and predict attitudes towards homosexuals.