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Studies of minute fossils trapped in amber shed light on the chronology of species and allow us to track evolutionary, ecological and biospheric changes. However, the resolution of commonly used methods for imaging amber fossils is limited, and the quality of preservation is not the only reason for this limitation. We have used super-resolution confocal laser microscopy (Airyscan) to study fossil mites (Acari) from the Eocene and Cretaceous amber (35–100 Ma). Here we report our technical approaches and methodological findings, including the selection of mounting medium, clearing of specimens, the effects of hydration of fossils in amber and the advantages of super-resolution confocal microscopy in obtaining images with sub-micron detail. Our results show that modern microscopy, in combination with specific non-destructive methods of amber preparation, can facilitate achieving image resolutions that were previously hardly feasible in fossils.

Pushing the limits of optical resolution in the study of the tiniest fossil arthropods