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Image Gallery: Increasing Confocal Resolution Down to 140 nm – HyVolution Confocal Super-Resolution Reveals More Details in Crisp Images

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.

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HyVolution resolves structures of 140 nm. Specimen: single molecule DNA-origamis with 140 nm defined spacing. Left: Confocal image. Right: HyVolution image. Distance measured matches 140 nm (white arrow).
HyVolution resolves structures of 140 nm. Specimen: single molecule DNA-origamis with 140 nm defined spacing. Left: Confocal image. Right: HyVolution image. Distance measured matches 140 nm (white arrow).
Applying HyVolution delivers detail-rich images with high contrast. MitoTracker Red CMXRos (mitochondria, yellow), Alexa Fluor 488 (F-actin, green), DAPI (nuclei, turquois). FluoCells Prepared Slide #1, LifeTechnologies.
Applying HyVolution delivers detail-rich images with high contrast. MitoTracker Red CMXRos (mitochondria, yellow), Alexa Fluor 488 (F-actin, green), DAPI (nuclei, turquois). FluoCells Prepared Slide #1, LifeTechnologies.
These images and the previous slide: Courtesy of Scientific Volume Imaging (SVI), Hilversum, The Netherlands
These images and the previous slide: Courtesy of Scientific Volume Imaging (SVI), Hilversum, The Netherlands
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.
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.
Resolution improvement in z. Mitochondrial membranes labelled with TOM20-GFP. Sample courtesy Urs Ziegler, ZMB, University of Zurich, Switzerland.
Resolution improvement in z. Mitochondrial membranes labelled with TOM20-GFP. Sample courtesy Urs Ziegler, ZMB, University of Zurich, Switzerland.
Paramecium, basal body (centriole-like structure at the base of flagella comprised of a 9*2+2 arrangement of microtubules). Deconvolution reveals the ring-like structure.
Paramecium, basal body (centriole-like structure at the base of flagella comprised of a 9*2+2 arrangement of microtubules). Deconvolution reveals the ring-like structure.
Leaf epidermal cells of Nicotiana Benthamiana after agrobacterium-mediated transient expression of P355-GFP. Courtesy of Mreille Chabaud, Toulouse.
Leaf epidermal cells of Nicotiana Benthamiana after agrobacterium-mediated transient expression of P355-GFP. Courtesy of Mreille Chabaud, Toulouse.
Paramecium aurelia. Epiplasmin (GFP, green), beta-tubulin (Alexa 568, red). Courtesy of Anne Aubusson-Fleury, CNRS, Gif-sur-Yvette, France.
Paramecium aurelia. Epiplasmin (GFP, green), beta-tubulin (Alexa 568, red). Courtesy of Anne Aubusson-Fleury, CNRS, Gif-sur-Yvette, France.
Applying HyVolution delivers detail-rich images with high contrast. MitoTracker Red CMXRos (mitochondria, yellow), Alexa Fluor 488 (F-actin, green), DAPI (nuclei, turquois). FluoCells Prepared Slide #1, LifeTechnologies.
Applying HyVolution delivers detail-rich images with high contrast. MitoTracker Red CMXRos (mitochondria, yellow), Alexa Fluor 488 (F-actin, green), DAPI (nuclei, turquois). FluoCells Prepared Slide #1, LifeTechnologies.
Increase in resolution and contrast by HyVolution imaging.
Increase in resolution and contrast by HyVolution imaging.
Endoplasmatic reticulum made clear: Medicago truncatula roots after agrobacterium-mediated transient expression of endoplasmatic reticulum-GFP. Courtesy of Mireille Chabaud, Toulouse.
Endoplasmatic reticulum made clear: Medicago truncatula roots after agrobacterium-mediated transient expression of endoplasmatic reticulum-GFP. Courtesy of Mireille Chabaud, Toulouse.
HyVolution alone does not reveal all details: Synaptonema complex. The true nature of the double strand can only be seen with STED super-resolution below 100 nm.
HyVolution alone does not reveal all details: Synaptonema complex. The true nature of the double strand can only be seen with STED super-resolution below 100 nm.
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.
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.

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.