Gliomas encompass a wide spectrum of various histopathological entities with different management strategies and associated prognoses. In many cases, initial biopsy of the brain lesion is required, since definitive diagnosis forms the foundation for treatment decision-making. Tissue sampling can be attained during stereotactic, open, or endoscopic procedures and, overall, provides >90% diagnostic yield, while it may be significantly lower (60-70%) in small (<1 cm3) and/or heterogeneous lesions. In the majority of the modern series, the morbidity rates do not exceed 2.5%. In experienced hands biopsy can be safely attained in any regions of the brain, including eloquent cortex, deep-seated structures, and the brainstem.