Serial synchrotron crystallography (SSX) is an emerging technique that leverages the advantages of serial crystallography (SX) to determine the noncryogenic crystal structure of macromolecules while minimizing radiation damage. SX, particularly serial femtosecond crystallography (SFX), uses X-ray free electron lasers (XFELs) to expose crystals only once during data collection, eliminating the need for cryogenic conditions. This method overcomes the limitations of conventional macromolecular crystallography (MX), which is prone to radiation damage and requires cryogenic temperatures. SSX, on the other hand, utilizes synchrotron X-rays, which are more accessible and provide a broader range of timescales for time-resolved studies. The technique involves continuous delivery of multiple crystals to the X-ray interaction point, with data collected from each crystal merged to form a complete dataset. Key considerations for SSX include optimizing photon flux, using high-viscosity injectors or fixed-target scanning techniques, and minimizing background scattering. The experimental setup and data processing procedures for SSX are detailed, highlighting the importance of efficient data collection and processing to enhance structural insights in structural biology.Serial synchrotron crystallography (SSX) is an emerging technique that leverages the advantages of serial crystallography (SX) to determine the noncryogenic crystal structure of macromolecules while minimizing radiation damage. SX, particularly serial femtosecond crystallography (SFX), uses X-ray free electron lasers (XFELs) to expose crystals only once during data collection, eliminating the need for cryogenic conditions. This method overcomes the limitations of conventional macromolecular crystallography (MX), which is prone to radiation damage and requires cryogenic temperatures. SSX, on the other hand, utilizes synchrotron X-rays, which are more accessible and provide a broader range of timescales for time-resolved studies. The technique involves continuous delivery of multiple crystals to the X-ray interaction point, with data collected from each crystal merged to form a complete dataset. Key considerations for SSX include optimizing photon flux, using high-viscosity injectors or fixed-target scanning techniques, and minimizing background scattering. The experimental setup and data processing procedures for SSX are detailed, highlighting the importance of efficient data collection and processing to enhance structural insights in structural biology.