Increasing use of Information and Communication Technology (ICT) for different purposes is evident. However, many Biology teachers and students hardly incorporate ICT into their instructional activities. Considering the major role of teachers and students in the classroom, especially in the use of ICT innovations, their perceptions must be considered as major factor in integrating ICT for classroom purposes. Therefore, this study investigated stakeholders’ perception of ICT use for Biology instruction in Oyo State. The objectives of the study were to:(i) examine the utilisation of ICT for teaching Biology in secondary schools; (ii)examine the perceived usefulness of ICT for teaching and learning by Biology teachers and students; (iii) determine the perceived ease of use of ICT for teaching and learning among Biology teachers and students; (iv) examine the gender differences in Biology teachers and students’ perceived usefulness of ICT for teaching and learning; and (v)investigate the gender difference on Biology teachers and students’ perceived ease of use of ICT for teaching and learning.rnA quantitative research design of the survey type was employed. The sample size included multistage sampled 45 biology teachers and 2578 biology students across nine Local Government Areas (LGAs) in Oyo State, Nigeria: nine LGAs were simple randomly sampled across three Senatorial Districts; and purposive sampling technique was used in selecting all the biology teachers, while, Research Advisor was consulted to select 2578biology students. Data was collected through a validated researcher-designed questionnaire with a reliability value of 0.92. Percentage (<50%>), mean and rank order (<2.50>) were employed to answer the research questions, while, hypotheses were tested using t-test and Analysis of Variance at 0.05 level of significance.rnThe findings of the study were that:rni. Biology teachers in Oyo State utilised ICT for teaching in secondary schools (54.7%>50%);rnii. the perceived usefulness of ICT for teaching and learning biology by biology teachers and students was positive in secondary schools in Oyo state (teachers: X=3.08; and students: X=2.68);rniii. the perceived ease of use of ICT for teaching and learning biology by biology teachers and students was positive in secondary schools in Oyo state (teachers: X=2.89; and students: X=2.68);rniv. there was no significant difference between male and female biology students’ perceived ease of use of ICT for learning in secondary schools in Oyo state; andrnv. there was no significant difference between male and female biology teachers’ perceived ease of use of ICT for teaching in secondary schools in Oyo state.rnThe study concluded that the effectiveness of ICT was determined not by its mere presence in the classroom but by teachers and learners’ use, perceived ease of use and perceived usefulness of the ICT. This implies that teachers’ and learners’ role is really critical as they are the ones who have to decide whether or not to use the available ICT resources in schools. This study recommended that ICT-based biology instructional processes should be made open, accessible and interactive to both the teachers and students.