Microbialites are sedimentary deposits associated with microbial mat communities and are thought to be evidence of some of the oldest life on Earth. Despite extensive studies of such deposits, little is known about the role of microorganisms in their formation. In addition, unambiguous criteria proving their biogenicity have yet to be established. In this study, we characterize modern calcareous microbialites from the alkaline Lake Van, Turkey, at the nanometer scale by combining x-ray and electron microscopies. We describe a simple way to locate microorganisms entombed in calcium carbonate precipitates by probing aromatic carbon functional groups and peptide bonds. Near-edge x-ray absorption fine structure spectra at the C and N K-edges provide unique signatures for microbes. Aragonite crystals, which range in size from 30 to 100 nm, comprise the largest part of the microbialites. These crystals are surrounded by a 10-nm-thick amorphous calcium carbonate layer containing organic molecules and are embedded in an organic matrix, likely consisting of polysaccharides, which helps explain the unusual sizes and shapes of these crystals. These results provide biosignatures for these deposits and suggest that microbial organisms significantly impacted the mineralogy of Lake Van carbonates.aragonite ͉ biosignature ͉ biomineralization ͉ spectromicroscopy L ake Van (eastern Anatolia, Turkey) is the largest soda lake on Earth, with a pH of 9.7-9.8 and a salinity of 21.7‰ (1). It harbors the largest known living microbialites, which are structures resulting from precipitation of aragonite at sites where calcium-rich groundwater seeps into the alkaline lake water (1, 2) and are associated with a wide diversity of microorganisms (3). Lake Van microbialites have a fine-grained micritic texture similar to most carbonate microbialites (4, 5) and consist of 30-to 100-nm-sized aragonite crystals (2, 3), which have morphologies that resemble bacteria-like forms (2). Some authors have suggested the possibility that nanospheres in microbialites could represent very small, entombed bacteria or ''nanobacteria'' (4, 5). However, the real nature of these carbonates, as well as their relationship to the microorganisms detected in the microbialites, remain unresolved.Because Lake Van is highly oversaturated with aragonite (1), the role of microorganisms in aragonite precipitation can be questioned, and the observed presence of microbes in these structures could simply result from passive trapping during mineral precipitation. This question is not new and has been raised systematically since the earliest studies of microbialites to the most recent ones (e.g., refs. 6-9). Some studies have demonstrated that microbes can actively mediate carbonate, in particular dolomite, precipitation (10). However, if passive trapping is operative, features suggesting that discrimination between microbially generated and purely abiotic precipitates is possible may be illusory. For example, the biogenicity of ancient stromatolites (i.e., laminated microbiali...