Functional annotations of the genome require a deep insight into signaling networks. This often needs sub 200 nm resolution for imaging of multiple stained specimens. Super-sensitive detection with low noise plus Huygens deconvolution is the key to resolving structures down to 140 nm with a confocal microscope. The result is crisp images full of contrast revealing details, which cannot be seen with confocal microscopy. This method can also be used for imaging with multiple colors simultaneously and in living cells.
The excellent signal-to-noise ratio of a photon counting hybrid detector enables to detect fluorescence from weakly emitting samples virtually without background noise. A high signal-to-noise ratio is a prerequisite for excellent deconvolution, as the background noise usually generated by PMTs is likely to produce more artifacts. Subsequent deconvolution reliably separates spots on single molecules like DNA origamis, which serve as nanorulers, with 140 nm defined spacing.
The possibilities of HyVolution confocal super-resolution can be explored in this image gallery.
HyVolution in multicolor live cells. All three colors were acquired simultaneously (no sequential scanning) in 5D. HeLa Kyoto cells. Golgi-GFP (green), GPI-YFP (yellow), H2B-mCherry (white). Sample courtesy of Sabine Reither, EMBL Heidelberg, Germany.