The Ozren ophiolite complex (OOC) of the Dinaridic Ophiolite Belt is one of the six ophiolite complexes in Bosnia and Herzegovina. This paper deals with the mineral chemistry of amphiboles determined by electron probe micro-analysis and micro-Raman spectroscopy. The detected amphibole generations and types in mafic, ultramafic, and metamorphic rocks suggest a polystage evolution and are therefore useful petrogenetic indicators of the investigated OOC. Most gabbroic rocks and dolerites contain primary magmatic amphibole1 (magnesio-hornblende to pargasite, occasionally hastingsite) and prismatic to needle-like aggregates of late magmatic amphibole2 (magnesio-hornblende), while plagiogranite contains ferri-winchite and ferro-ferri-winchite as primary magmatic amphibole. Post-magmatic amphiboles were detected in dolerites, troctolites, and lesser in peridotites. The Na-(Ti)-rich amphibole3 (ferri-winchite and ferro-ferri-winchite to katophorite and ferri-katophorite) with amphibole4 (grunerite) rim formed along the grain boundaries of clinopyroxene, amphibole1, and plagioclase in dolerites. A part of these amphiboles grows into amphibole1, 2. Kaersutite to ferri-kaersutite, associated with phlogopite, occur in troctolites and dunites, while Mhbl was detected in harzburgite. The ultramafic rocks (lherzolites, harzburgites, and dunites) and the gabbroic layer are crosscut by clinopyroxene–plagioclase gabbroic and clinopyroxene–plagioclase–amphibole gabbro–dolerite dykes, suggesting ‘dry’ and ‘hydrated’ percolating melts generated in inferred subridge and supra-subduction settings, respectively. The amphibole3 and 4 in gabbros and dolerites and similar amphibole types in ultramafic rocks could be related to inferred arc-type basaltic and plagiogranitic percolating melts and fluids. Low-Al amphibole5 (tremolite and actinolite) and associated chlorite, albite, and clinozoisite represent the ocean-floor alterations in mafic rocks. Amphibole6 (magnesio-hornblende to pargasite) was identified in metamorphic sole amphibolites. Micro-Raman spectroscopy provided typical Raman spectra for the studied amphiboles, highlighting distinct features such as bands related to CMg content, CFe3+ presence, TO4 ring-breathing mode, TiO6 stretching mode, presence > 0.3 apfu of CTi, and TO4 stretching indicating CFe2+ in the structure. Applied amphibole geothermobarometry revealed the formation P–T conditions of amphibole (Amp)1 (avg. 863 °C at 0.23 GPa), Amp2 (avg. 747 °C at 0.17 GPa), Amp in the mantle rocks (avg. 853 °C at 0.64 GPa), Amp5 (avg. 349 °C at 0.03 GPa), and Amp6 (avg. 694 °C at 0.46 GPa).