The aim of this study is to identify the relationship between psychopathological symptoms, coping strategies, and the sexual orientation of Brazilian nursing professionals in the context of the COVID-19 pandemic. A cross-sectional and analytical study with 1737 nurses, midwives, nursing assistants, and technicians, working at different levels of health care, direct assistance and/or administrative/managerial assistance, and linked to teaching and research. These included socio-demographic data, Psychopathological Symptom Assessment Scale, and coping strategies inventory. For data analysis, descriptive statistics and Wilcoxon, Mann–Whitney, and Kruskal Wallis tests were used, with a significance level of 0.05. Statistically significant differences were evidenced between psychoticism; obsessiveness-compulsivity; somatization and anxiety, indicating mental suffering. Bisexual professionals showed greater severity of psychopathological symptoms. As for the coping strategy, there is a statistically significant difference in the mean scores of Withdrawal, Responsibility, Escape from Fear, and Self-Control, according to sexual orientation. The factors Withdrawal, Self-control, and Responsibility had higher means in bisexuals (
p
< 0.05) and lower means in confrontation (
p
> 0.05). In homosexuals, the highest mean scores were for the problem solving and social support factors (
p
> 0.05) and lowest in the Withdrawal factor (
p
< 0.05). Among heterosexuals, the highest mean score was in confrontation (
p
> 0.05) and the lowest in avoidance (
p
< 0.05). This study reinforces the importance of reflecting on the life context and work settings of nursing professionals, especially bisexual women, through the influence of the effects of the Covid-19 pandemic on their mental health. The data can guide actions that minimize the impacts on mental health, in addition to recognizing, based on the findings, a collective complaint from the professional category to the health authorities and competent bodies, as a way of explaining the context experienced in the locus of their work.