A real-time imaging acquisition and analysis system is designed to investigate thin film growth on transparent substrates using fluorescence microscopy. And eight growth parameters are proposed based on film growth morphological feature. Besides, using home-made experimental system, we analyze the p-6P nanofiber growth on the mica substrate dynamically, revealing the quasi-one-dimensional growth mode of the para-sexipheny (p-6P) molecules on the mica substrate. As an important imaging technique for monitoring thin film growth, the system is expected to play an active role in understanding micro-structural properties of thin film and the interactions between film and substrate.