This unit describes methods for growing and analyzing static biofilms, which are useful for studying early stages of biofilm formation, including initial adherence and microcolony formation. Static biofilm assays are simple, require minimal equipment, and allow for various analyses such as microscopy, staining, and viability counts. These assays are suitable for both small and large-scale studies and can be adapted to various conditions. They are particularly useful for identifying signals that modulate the transition from a planktonic to a biofilm lifestyle. However, they may have limitations in generating mature biofilms due to the lack of continuous nutrient supply and aeration. Continuous-flow and chemostat systems are described in other units for addressing these limitations. The unit presents four basic protocols for static biofilm growth and analysis. The microtiter plate biofilm assay is a high-throughput method for assessing bacterial attachment by staining adherent biomass. The air-liquid interface (ALI) assay allows direct microscopic observation of live attached microbes. A colony-based biofilm system is useful for monitoring cell death in biofilms treated with antimicrobial agents. The Kadouri system is a "low-flow" system that bridges static assays and continuous-flow systems. The microtiter plate assay involves growing bacteria in microtiter dishes, washing away planktonic cells, and staining adherent biomass. The ALI assay involves tilting a plate to create an air-liquid interface for microscopic analysis. The colony biofilm assay uses a semipermeable membrane on an agar plate to grow biofilms, allowing for nutrient replenishment. The Kadouri system uses a peristaltic pump to continuously supply and remove medium, enabling the growth of mature biofilms. These assays are valuable for studying biofilm formation, antibiotic resistance, and the effects of various environmental factors. They can be adapted for genetic screens, small molecule screening, and real-time monitoring of biofilm dynamics. The unit also includes troubleshooting tips and critical parameters for successful execution of these assays.This unit describes methods for growing and analyzing static biofilms, which are useful for studying early stages of biofilm formation, including initial adherence and microcolony formation. Static biofilm assays are simple, require minimal equipment, and allow for various analyses such as microscopy, staining, and viability counts. These assays are suitable for both small and large-scale studies and can be adapted to various conditions. They are particularly useful for identifying signals that modulate the transition from a planktonic to a biofilm lifestyle. However, they may have limitations in generating mature biofilms due to the lack of continuous nutrient supply and aeration. Continuous-flow and chemostat systems are described in other units for addressing these limitations. The unit presents four basic protocols for static biofilm growth and analysis. The microtiter plate biofilm assay is a high-throughput method for assessing bacterial attachment by staining adherent biomass. The air-liquid interface (ALI) assay allows direct microscopic observation of live attached microbes. A colony-based biofilm system is useful for monitoring cell death in biofilms treated with antimicrobial agents. The Kadouri system is a "low-flow" system that bridges static assays and continuous-flow systems. The microtiter plate assay involves growing bacteria in microtiter dishes, washing away planktonic cells, and staining adherent biomass. The ALI assay involves tilting a plate to create an air-liquid interface for microscopic analysis. The colony biofilm assay uses a semipermeable membrane on an agar plate to grow biofilms, allowing for nutrient replenishment. The Kadouri system uses a peristaltic pump to continuously supply and remove medium, enabling the growth of mature biofilms. These assays are valuable for studying biofilm formation, antibiotic resistance, and the effects of various environmental factors. They can be adapted for genetic screens, small molecule screening, and real-time monitoring of biofilm dynamics. The unit also includes troubleshooting tips and critical parameters for successful execution of these assays.