Polystyrene (PS) is the most commonly used material in cell culture devices, such as Petri dishes, culture flasks and well plates. Micropatterning of cell culture substrates can significantly affect cell-material interactions leading to an increasing interest in the fabrication of topographically micro-structured PS surfaces. However, the high stiffness combined with brittleness of PS (elastic modulus 3–3.5 GPa) makes high-quality patterning into PS difficult when standard hard molds, e.g. silicon and nickel, are used as templates. A new and robust scheme for easy processing of large-area high-density micro-patterning into PS film is established using nanoimprinting lithography and standard hot embossing techniques. Including an extra step through an intermediate PDMS mold alone does not result in faithful replication of the large area, high-density micropattern into PS. Here, we developed an approach using an additional intermediate mold out of OrmoStamp, which allows for high-quality and large-area micro-patterning into PS. OrmoStamp was originally developed for UV nanoimprint applications; this work demonstrates for the first time that OrmoStamp is a highly adequate material for micro-patterning of PS through hot embossing. Our proposed processing method achieves high-quality replication of micropatterns in PS, incorporating features with high aspect ratio (4:1, height:width), high density, and over a large pattern area. The proposed scheme can easily be adapted for other large-area and high-density micropatterns of PS, as well as other stiff and brittle polymers.