A precise and rapid mapping protocol for correlative light and electron microscopy of small invertebrate organisms

• DOI: 10.1042/bc20090096
• Publication Date: 2009-09-22T09:51:09Z
• Authors: Irina Kolotuev; Yannick Schwab; Michel Labouesse
• Keywords: Correlative; Multicellular organism; Immunoelectron microscopy

CLEM (correlative live cell and electron microscopy) seeks to bridge the data acquired with different imaging strategies, typically between light microscopy microscopy. It has been successfully applied in cultures, although its use multicellular systems is hampered by difficulties locating ROI (region of interest).We developed a technique that enables easy processing small model animals adequate both for morphology immunoelectron-microscopic specimen preparations. While this method initially Caenorhabditis elegans samples, we found it works equally well Drosophila samples. handling observation single any complex genotype real time, fixation high-pressure freezing flat embedding. Our major improvement development precise mapping system considerably simplifies speeds up retrospective location within 1 mum distance. This can be used when correlative required, as facilitate treatment non-correlative TEM procedures. improvements open possibility treat statistically significant numbers processed electron-microscopic protocols, making them more accessible wider range researchers.We believe will contribute studies models time-demanding procedure preparation kind TEM.