When studying subcellular detail both, resolution and sensitivity of a confocal microscope can become limiting. Different approaches to improve the resolution of a confocal microscope are available, but they are constantly challenging the sensitivity of the system.
In this presentation we demonstrate how we improve the resolution of the Leica TCS SP8 confocal microscope using super-sensitive HyD hybrid detectors and Huygens deconvolution in our new HyVolution system. Compared to the usual diffraction limit around 240 nm, we achieve sub-diffraction limited resolution of 140 nm laterally and an axial resolution increase by a factor of 2.
This is possible, because of the high signal-to-noise ratio of the HyD detector. Being a photon-counting detector Leica HyD circumvents noise amplification associated with traditional intensity averaging. Its superior signal-to-noise ratio compared to (GaAsP) photomultipliers thus translate into higher resolution when combined with state of the art deconvolution.
Even single molecules such as DNA origamis can be reliably resolved as demonstrated using a 140 nm nanoruler.
The main advantage of using HyVolution over other techniques addressing this resolution domain is that you can freely choose acquire multiple colors simultaneously at full speed. This makes it ideal for fixed specimen and live cell imaging alike.