Microbial, specifically fungal and bacterial infections are the prime cause of diseases in humans, animals, and plants. Technological advancements with molecular protocols have undoubtedly broadened the research areas against global microbial infections. High antifungal and antibacterial drugs usage with resistance development, host-specific toxicity, mutational alterations, and several other factors bring forth the urge for new antimicrobial agents. New protocols related to antimicrobial activities and developing novel drugs against severe infections are highly recommended in future research areas. Dilution, diffusion, bioautography protocols, and several others are frequently used to determine the antimicrobial activities. The aim of this review is to highlight and assemble the information regarding the various anti-fungal activity techniques. Additionally, their principle, methodology, advantages, and disadvantages, with information regarding solvents and microbial organisms tested, has also been reviewed.