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Is confocal microscopy obsolete?

Super-resolution microscopy is at the forefront of biomedical science today, but does it supersede confocal microscopy?

Of course not! In fact, the opposite is true – good confocal imaging is the ideal base from which to blast off on a journey beyond the diffraction limit.

Confocal Microscopy alone provides better lateral resolution than the Abbe limit, if operated in the appropriate regime. And only confocal microscopes that perform at high sensitivity and low noise can benefit from these confocal-inherent resolution improvements. In combination with sensible and professional image reconstitution algorithms, it is possible to achieve resolution of 140 nm resolution and beyond, at high speed and multiple channels simultaneously, all without having to tinker endlessly with the optical setup.

* See the difference: Two color confocal versus HyVolution Super-Resolution image of Paramecium tetraurelia. Green: Basal bodies (AF488), red: Epiplasmin (AF568). Sample courtesy of Anne Aubusson-Fleury, Institute for Integrative Biology of the Cell (I2BC), French National Centre for Scientific Research (CNRS), Gif sur Yvette, France

Find out

what HyVolution Super-Resolution can do for your research


Transform your confocal microscope into a super-resolution microscope

Leica’s HyVolution solution transforms a confocal microscope into a super-resolution microscope. Teaming ingenious inventions with smart implementation has resulted in a confocal microscope with continuously tunable filter functions, the Leica TCS SP8. It seamlessly adapts to any dye or dye combination you may want to employ. As a second pillar, the tunable elements provide maximal transmission. This prevents loss of your precious emission, whilst enabling very thin sectioning conditions. After all, optical sectioning is the main reason for buying a confocal microscope.

Compare the results: 3 channel simultaneous acquisition of COS7 cells with HyVolution super-resolved (right) versus confocal (left). Green: Mitochondria (Oregon Green 488), red: Actin (SiR), white: Tubulin (TMR). Sample courtesy of Dr. Jana Doehner, Center of Microscopy and Image Analysis, University of Zurich, Switzerland.

Don't overload your optical beam path!

High sensitivity and thin optical sections then merge with scientific image reconstitution to result in super-resolved images with high contrast and very low noise. That is exactly what you need.

HyVolution does not overload the optical beam path with additional elements, no fibers are required to transmit the emission and no optics are needed to adapt the diffraction pattern. All channels can operate in parallel in super-resolution mode, at all magnifications and wavelengths independently, including UV- and IR-excitation. And it is available for non-descanning detector signals too.

If you want to image living samples speed is what matters. The Leica TCS SP8 offers truly parallel channel recording, ensuring no decrease in imaging speed, whatever configuration you choose.

HyVolution Super-Resolution impressions

Rat cortical neurons (prim. Culture)
Blue: Nuclei (Dapi), red: Nestin (Cy2), yellow: DCX (Cy3, green: βIII-tubulin (cy5).
Rat cortical neurons (prim. Culture) acquired with HyVolution Super-Resolution. Blue: Nuclei (Dapi), red: Nestin (Cy2), yellow: DCX (Cy3, green: βIII-tubulin (cy5).
Paramecium tetraurelia
Green: Epiplasmin (GFP), red: β-Tubulin (Alexa 568). Sample courtesy of Anne Aubusson-Fleury, Institute for Integrative Biology of the Cell (I2BC), French National Centre for Scientific Research (CNRS), Gif sur Yvette, France.
Paramecium tetraurelia acquired with HyVolution Super-Resolution. Green: Epiplasmin (GFP), red: β-Tubulin (Alexa 568). Sample courtesy of Anne Aubusson-Fleury, Institute for Integrative Biology of the Cell (I2BC), French National Centre for Scientific Re
Mouse testis
Green: Synaptonemal Complex (Alexa 488), blue: Nuclei (DAPI), red: Protein X (Cy3). Courtesy of Yu Katsuyama and Noriko Osumi, Tohoku University, Miyagi, Japan.
Mouse testis acquired with HyVolution Super-Resolution. Green: Synaptonemal Complex (Alexa 488), blue: Nuclei (DAPI), red: Protein X (Cy3). Courtesy of Yu Katsuyama and Noriko Osumi, Tohoku University, Miyagi, Japan.