Gliosarcoma with areas of primitive neuroepithelial differentiation (GSPNED) is an extremely rare neoplasm. A case is presented here in which squash smears of a left temporal lobe tumor in a 76-year-old male demonstrated two distinct and easily recognizable cellular populations, i.e., densely hyperchromatic cells of a primitive nature in a fibrillary background and pleomorphic spindle-shaped cells. Occasional pseudo-rosette formations and nuclear cannibalism suggestive of neuroendocrine differentiation were also found. A cytologic diagnosis of a malignant tumor was suggested, and histochemical and immunohistochemical studies were conducted on formalin-fixed, paraffin-embedded material. Reticulin stain highlighted increased intercellular collagen and reticulin deposition within the spindled regions, whereas nodules with primitive cells were reticulin-poor. There was a diffuse and strong reactivity to neuron specific enolase, synaptophysin and CD56 immunostains. A stain for glial fibrillary acidic protein and S-100 protein demonstrated a subset of tumor cells including elongated cytoplasmic processes. The spindled component was positive for vimentin and smooth muscle actin, whereas the primitive-appearing tumor cells were negative. The diagnosis of GSPNED was confirmed based on cytopathologic, histopathological and immunohistochemical results. The cytomorphologic features of this distinctive tumor are illustrated, and the adjunctival value of squash smears for frozen-section diagnosis is also discussed. This is the first presentation of a cytopathologic analysis that provides an important clue to an accurate diagnosis of GSPNED.