OBJECTIVE To describe clinical experience in the diagnosis of intraocular lymphoma by fine-needle aspiration biopsy (FNAB) in patients with one or more discrete intraocular infiltrative lesions and limited or absent intravitreal tumour cells. METHODS Retrospective descriptive analysis of patients who underwent intraocular FNAB of a solid retinal, subretinal pigment epithelial or uveal tumour that proved to be a malignant lymphoma. RESULTS After exclusions, our study group consisted of seven patients, each of whom had one or more discrete intraocular infiltrative lesions and limited or absent intravitreal tumour cells and underwent a diagnostic intraocular FNAB that confirmed malignant intraocular lymphoma cytopathologically. These included three patients with one or more geographic yellow subretinal pigment epithelial infiltrates and one patient each with a prominent nodular white subretinal pigment epithelial tumour, a rapidly developing solid placoid choroidal mass, a haemorrhagic retinal infiltrative lesion and an infiltrative iris tumour, respectively. A prominent feature of virtually all aspirates was a large proportion of degenerated lymphoid cells in the background. Cytologically intact tumour cells ranged from relatively homogeneous small round cells with large nucleus to cytoplasm ratio to pleomorphic large cells with irregular knob-like nuclear protrusions. Immunocytochemical stains for lymphoid markers were helpful in confirming the pathological diagnosis of lymphoma in the five patients in whom this testing was performed. CONCLUSIONS FNAB was a useful diagnostic tool in the described subgroup of patients with suspected intraocular lymphoma. FNAB should be considered as a diagnostic option in selected patients with suspected intraocular lymphoma, especially if there are few or no vitreous cells.