Single Molecule Imaging - Applications To Cellular Processes

Using high-affinity antibody-based probes, multimerized epitope tags, and single molecule microscopy, we are able to image single RNA translation dynamics in living cells and quantify the emergence of nascent protein.

This technology will be described as well as a two color extension, useful for comparing translation rates between two different parts of a single open reading frame (ORF) or two different ORFs. Using information from the correlations of fluorescence fluctuations, we can accurately quantify single mRNA translation elongation rates in both tagged and untagged portions of ORFs. By transiently loading probes and reporter DNA into cells in a combinatorial fashion, multiplexed imaging of gene expression is possible.

This webinar will introduce the new iXon Life EMCCD cameras and provide performance comparison with the well-known iXon Ultra cameras. Both camera series are the best-in-class detectors for sensitivity allowing short exposure times and thus reduced photo-toxicity and photo-bleaching. Used in ‘crop mode’, EMCCDs can image at high frame rate with e.g. 93 fps for a sub-array of 512 x 512 pixels.