AbstractShared resource laboratories/core facilities (SRLs) are centralized platforms that house and provide access to complex and expensive research equipment. Due to the highly complex nature of the instrumentation they support, SRLs have special environmental requirements for their laboratory space. Here, we describe the planning and establishment of a large light microscopy SRL, with a special focus on room layout, custom‐designed air conditioning and vibration, which can also be adapted to proteomics, genomics, and flow or mass cytometry SRLs.