In Situ Real-Time Study on Dynamics of Microbially Induced Calcium Carbonate Precipitation at a Single-Cell Level

Ureolytic microbially induced calcium carbonate precipitation (MICP) is a promising green technique for addressing a variety of environmental and architectural concerns. However, the dynamics of MICP especially at the microscopic level remains relatively unexplored. In this work, by applying a bacterial tracking technique, the growth dynamics of micrometer-sized calcium carbonate precipitates induced by Sporosarcina pasteurii were studied at a single-cell resolution. The growth of micrometer-scale precipitates and the occurrence and dissolution of many unstable submicrometer calcium carbonate particles were observed in the precipitation process. More interestingly, we observed that micrometer-sized precipitated crystals did not grow on negatively charged cell surfaces nor on other tested polystyrene microspheres with different negatively charged surface modifications, indicating that a negatively charged surface was not a sufficient property for nucleating the growth of precipitates in the MICP process under the conditions used in this study. Our observations imply that the frequently cited model of bacterial cell surfaces as nucleation sites for precipitates during MICP is oversimplified. In addition, additional growth of calcium carbonates was observed on old precipitates collected from previous runs. The presence of bacterial cells was also shown to affect both morphologies and crystalline structures of precipitates, and both calcite and vaterite precipitates were found when cells physically coexisted with precipitates. This study provides new insights into the regulation of MICP through dynamic control of precipitation.