Staining for Granulosa cells and Immunofluorescence staining for FSH receptor. (A) Aggregated granulosa cells stained with Papanicolaou (20x magnification). Bar = 20 μm. Localisation of FSH receptor (FSHR) in human granulosa cells (GC) after six days in culture, Immunofluorescence FSHR staining with monoclonal mouse anti-human FSHR (primary), chicken anti-mouse (Alexa, secondary), counterstained with DAPI for nuclei. (B) GCs in control culture (without FSH) showing cytoplasmic vesicles; (C) GC culture stimulated with FSH 0.1 IU/mL showing outgrowth with fibroblast-like morphology; (D) GC culture stimulated with FSH 1 IU/mL showing an intracellular signal for FSHR which could indicate de novo production (40x magnification). Bar = 40 μm.