In the imaging sciences we are often interested in the precise geometric localisation of biological structures. Through using optical microscopy techniques we are able to visualise fluorescently tagged or stained biological structures and infer their geometric localisation. An artefact of this visualisation process is often out-of-focus light which can reduce our ability to segment and localise specific structures of interest. Image deconvolution is often applied to compensate for the presence of out-of-focus light and tools such as those provided by Huygens specialise in this process. Used with care, Huygens can improve the clarity of microscopy images where there is significant out-of-focus light allowing for more discrete analysis to be applied. Deconvolution is most often applied to wide-field images however it is also applicable for confocal microscopy and super-resolution techniques where some out-of-focus light is still present. For more documentation on Huygens the following website is recommended: http://www.svi.nl/HuygensSoftware. For more notes and advice about deconvolution please refer to:. For more information relating to this or for an introduction to the system please contact Dominic Waithe.
An artefact of glass optics is that their ability to bend light is wavelength dependent. The result of this effect is that light of differing wavelength, but which is originating from the same source, can appear shifted, in a slightly different localisation. This artefact known as chromatic shift aberration, is very apparent when localising objects in the z-plane. This artefact can be detrimental for establishing localisation or colocalisation characteristics across different wavelengths. Huygens software can also be used to correct for chromatic shift . A guide for correcting for chromatic shift using Huygens can be found at: http://www.svi.nl/ChromaticShiftCorrector
Based on our CBRG servers we have two Huygens licences:
- On the Klyn server we have a medium server option for Huygens which allows 5-users to simultaneously perform wide-field deconvolution. Image files produced on wide-field systems including the Wolfson Imaging Centre DeltaVision microscope are suitable for processing using this server. This server is rated as the 2nd fastest Huygens implementation in the world.
- Through STimulated Emission Depletion (STED) microscopy it is possible to achieve a theoretically unlimited resolution. In practise however the resolution of STED is limited by noise, as a consequence of the small number of photons emerging from biological preparations at super-resolution. It has been found that noise in STED images can be reduced using Huygens deconvolution and the resolution further optimised . On the Deva server we have a Huygens server for STED deconvolution. This server can be used by one user at a time. For more information
If users wish to use these facilities they must have a CBRG account: http://www.molbiol.ox.ac.uk/CBRG_accounts.shtml
For more information relating to any of features described above, please contact Dominic Waithe.
 Measuring and interpreting point spread functions to determine confocal microscope resolution and ensure quality control. Cole RW, Jinadasa T & Brown CM. Nat Protoc. 2011 6: 1929-41.
 A workingperson's guide to deconvolution in light microscopy. Wallace W, Schaefer LH, Swedlow JR. Biotechniques. 2001 31:1076-8, 1080, 1082 passim.
 Huygens STED Deconvolution Increases Signal-to-Noise and Image Resolution towards 22 nm. Schoonderwoert V, Dijkstra R, Luckinavicius G, Kobler O and van der Voort H. Microscopy Today 2013 21: 38-44.