In recent years there has been a growing interest in researching and developing new antimicrobial agents from various source to combat microbial resistance. Therefore, a greater attention has been paid to the antimicrobial activity screening and evaluating methods. Several bioassays such as disk diffusion, well diffusion are well known and commonly used, but other such as flow cytofluorometric and bioluminescent are not widely used because they require specified equipment’s and further evaluation for reproducibility and standardization, even if they can provide rapid results of the antimicrobial agents effects and a better understanding of There impacts on the viability and cell damage inflicted to the tested microorganism.in this review article, and exhaustive list of in vitro antimicrobial sustainability testing method and detailed information on their advantages and limitations are reported.