Background: E-learning is a powerful tool that transforms the traditional learning method to strengthen the capacity of teaching and learning. Detecting challenges of E-learning met by undergraduate nursing students could contribute to improving the quality of the learning environment in nursing education. Aim: evaluate university nursing students' Perception and challenges towards E-learning and recommend overcoming ways. Design: A mixed-methods triangulation design was used to collect both quantitative and qualitative data through a web-based form. Setting: The study was conducted at the Faculty of Nursing affiliated to Ain Shams University, Cairo, Egypt. Subjects: The convenient sampling technique composed of (340) university nursing students enrolled in the nursing program during the second semester of the academic year (2022-2023), in the second and fourth batches. Data collection tools: It consists of four parts: demographic characteristics, the perception of students toward E-learning, Challenges of E-learning, and the recommended overcoming ways. Results:  The majority of second and fourth academic years nursing students had positive Perception of the benefits of E-learning. while their Perception during attending e-learning for fourth-year nursing students were higher than second year. In addition, three-quarters of the fourth-year academic nursing students reported no challenges faced them during E-learning compared with two-thirds of the second year. Conclusion: there was a positive statistical correlation between overall Perception and challenges toward E-learning as reported by nursing students. Likewise, there was a statistically significant difference between the student's academic years and their overall Perception and challenges toward E-learning as reported by nursing students. Recommendations: Enhance Internet speed and make Internet packages cheaper or even free and supply e-learning tools and computer skills training for students.