Microbial mats, mainly dominated by filamentous algae Calothrix and Oscillatoria, are well developed in Tibetan hot springs. A great number of fossil microorganisms, which existed as algae lamination in thermal depositional cesium-bearing geyserite in this area, are identified as Calothrix and Oscillatoria through microexamination and culture experiments. These microbial mats show the ability to accumulate cesium from spring water to the extent of cesium concentration of 0.46–1.03% cell dry weight, 900 times higher than that in water, and capture large numbers of cesium-bearing opal grain. Silicon dioxide colloid in spring water replaces and fills with the organism and deposits on it to form algae laminated geyserite after dehydration and congelation. Cesium in the microbial mats and opal grain is then reserved in the geyserite. Eventually, cesium-bearing algae laminated geyserite is formed. Study on cesium distribution in geyserite also shows that cesium content in algae lamination, especially in heavily compacted algae lamination, is higher than in the opal layer. For geyserite with no algae lamination or other organism structure, which is generally formed in spring water with low silicon content, cesium accumulation and cesium-bearing opal grain assembled by the microbial mats are also indispensable. After the microbial mats accumulating cesium from spring water, silicon dioxide colloid poorly replaces and fills with the organism to form opal grain-bearing tremellose microbial mats. The shape and structure of the organisms are then destroyed, resulting in cesium-bearing geyserite with no algae lamination structure after dehydration and congelation. It is then concluded that microbial mats in the spring area contribute to the enrichment of cesium in the formation of cesium-bearing geyserite, and a biological genesis of the geyserite, besides of the physical and chemical genesis, is likely.