The special characteristics of ocean ecosystems, such as the high salinity and pressure, low temperature, and nutrition, makes marine microorganisms diverse in species, gene composition, and ecological functions. Recent advances in molecular biology techniques, together with the ongoing developments in bioinformatic and automatic technologies, have highlighted the scientific studies on marine microbial ecology, eliminating the total reliance on classical cultivation-based techniques. This review systematically summarizes the innovative aspects of a field investigation of marine microorganisms. It covers various cutting-edge sampling approaches employed in the field, highlighting the in situ high-fidelity sampling strategies with practical applications. Particular emphasis is placed on the molecular analyses for marine ecology based on recently developed omics technologies. The key technical issues and substantial contribution of the contemporary instrumental analyses are discussed accordingly. It provides references for the in situ surveys of complex biogeochemical processes from marine microorganisms to ecosystems, relying on the development of new technical concepts and scientific methodologies in field investigations.