Cellular replication and atomic force microscope imaging using a UV-Bioimprint technique

Replication and fixation techniques have been of considerable interest for imaging and analysis of biological cells since the introduction of electron and scanning probe microscopy. Although such tools as the atomic force microscope (AFM) permit in situ morphological studies at a magnitude of resolution beyond traditional optical microscopy, they are diffcult to operate and their resolution capabilities are rarely realized. We used a UV-Bioimprint replication technique to imprint a polymer layer onto cells attached to a substrate and rapidly cure to create an impression of cell topography. Replicas of chemically fixed and untreated cells analyzed by atomic force microscopy demonstrate nanometer resolution in the transfer of replicated features. UV-Bioimprint presents an improvement over techniques using heat-curable polymers as well as an alternative technique to the direct imaging of cells. The motivation for UV-Bioimprint is to effectively integrate scanning probe microscopy tools for imaging of cellular ultrastructure.