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Calcium-binding protein Iba-1 is currently believed to be identical to AIF-1 protein and is regarded as a selective marker for microglia. However, data on populations of cells able to express Iba-1/AIF-1 are contradictory. The aim of the present work was to identify rat brain cells expressing Iba-1 protein and to describe their functional characteristics. Immunocytochemical and confocal laser microscopy methods were used. Iba-1 protein was seen only in microglial cells, meningeal macrophages, supraependymal macrophages, and superficial and stromal cells of the vascular plexus, i.e., cells with phagocytic functions. Comparison of these data with results from other studies suggests that not all of the currently used antibodies to Iba-1/AIF-1 protein recognize the same product. This may be associated with alternative splicing of the Iba-1 mRNA.