Scanning electron microscopy (SEM) of insect and plant fossils in the lacustrine shales of the Eocene Florissant Formation of Florissant, Colorado, was used to investigate the mechanisms of fossil preservation. The fossiliferous Florissant “paper shales” are composed of thin laminae of diatomite that form couplets with alternating smectitic clay laminae. The millimeter-scale sedimentary couplets may preserve an episodic record of sedimentation and are interbedded with less frequent, coarser volcaniclastic layers. The insect and plant fossils are associated with biofilms of extracellular polymeric substances (EPS) secreted by diatoms. The preserved organisms are entangled in the diatom aggregates coated with the EPS biofilm. We suggest that decomposition of the organisms was arrested during sedimentation and burial by the protective nature of the mucus covering, the properties of which limited the actions of bacteria and grazers and may have enhanced fossilization. A novel contribution of the study is a demonstration that this mechanism of exceptional preservation is also common at other similar lacustrine fossil sites, as supported by a further SEM analysis of insect and plant fossils from other Cenozoic lake deposits formed in environments comparable to the Floris-sant Formation. The deposits include the Oligocene shale at Canyon Ferry, Montana; the Miocene Savage Canyon Formation, Stewart Valley, Nevada; and the Miocene Shanwang Beds of Shandong Province in northeast China. In addition, cultures of diatomaceous biofilms, grown in the laboratory display morphological features identical to those of the fossil diatomaceous biofilms. Our contribution indicates the significance of biofilms in fossil preservation at Florissant and other deposits.