Although neurogenesis has been well studied, its molecular mechanisms remain largely unknown due to the challenges posed by the complexity of the underlying processes. Whilst in vivo studies can be used to study neurogenesis, the inability to control confounding variables complicate findings. Therefore, the purpose of this study was to identify the markers of in vitro neural rosette formation and describe the formation of neural rosettes from pluripotent stem cells using immunofluorescence analysis. The protocol of stem cell cultivation and induction of neural rosette formation was tested. Following, two transcription factors, BRN2 and SOX2, were fluorescently labelled and cells were imaged over a period of eight days. It was identified that SOX2 and BRN2 are expressed during in vitro neural rosette formation. These results are concurrent with in vivo neurogenesis, which suggests that neural rosettes could be a suitable in vitro model for researching neural development. Given that mistakes can arise during neurogenesis, such as neural tube defects, developing robust models to understand the formation of the nervous system is important. Moving forward, a detailed molecular understanding of neural rosette formation has the potential to be used for targeting specific transcription factors to treat or prevent problematic neurogenesis.