In Life Sciences Education, the use of educational external representations (ERs) such as diagrams, models and animations are increasingly appearing in learning and teaching resources. However, their effectiveness is limited if learners experience learning difficulties due to lack of visual literacy and spatial ability skills to work with ERs. The study explored the level of visual literacy of 225 Grade 10 Life Sciences learners in cytology across six secondary schools in Pretoria, Gauteng. It was theorised that ERs need to be integrated in the Life Sciences curricula to develop learners’ visual literacy and spatial ability skills. With this theory, the study aimed to explore the visual literacy of Grade 10 Life Sciences learners and the influence of gender and school location on the visual literacy and spatial ability skills of the learners. Through a quantitative research method a Life Sciences visual literacy questionnaire and a spatial ability test were used to collect data. Collected data was analysed descriptively and inferentially through Statistical Package Social Sciences Version 23. The results showed that most Grade 10 Life Sciences learners lack average visual literacy skills. Furthermore, the results showed that gender doesn’t play a role on the learners’ performance in visual literacy skills as both genders performed equally in both tests, On the other hand, the results showed that the location of the school (urban, rural or township) has an effect on the learners’ performance in visual literacy skills. Teachers need to incorporate different ERs that would stimulate different senses and which will also enhance learners’ visual literacy and spatial ability skills in their lessons. A conclusion and some recommendations for future research are given.