Nikon Imaging Center at Hokkaido university

About deconvolution and Huygens

Huygens is powerful for Deconvolution tool!

Huygens supported by Solution Sysytemsis the software to operate deconvolution. Deconvolution is a mathmatically method for eliminating optical blur from out focus singal. Confocal microscope can only catch in focus sigals with two pinhole, but it is not completely. Then fluorescence signal have to contains from not only in focus signal but also out focus one. Therefore, operating to deconvolution, users enable to recover original signal and to make their images clealy.

Since deconvolution is neither change imaging data nor image enhancement, you have only to write down the detail of deconvolution (for example, methods and software) in your paper.

*Example of paper. (sorry, now under reconstruction.)

The details are shown at Solution Systems and SVI (Huygens Gallery). Please refer to it.

The strong point of Huygens

  • If you didn't acquire several frames for Z direction, you can process the deconvolution for single frames. And Huygens can also operate successive process, you can do for multi channel time lapse images at once.

  • As expected above, it is enalbe to improve the quality for images acquired not only by wide field fluorescence images (at Station-3) but also by confocal microscope (at Station-1, Station-4) or multi photon microscope.

  • Since Huygens equipped with 3D viewer, users can easily make some movies of 3D rendering models and their rotation / zooming up.

Attention for deconvolution

  • Users have to set some parameters exactly - EX and EM wavelength, information of objective and pinhole size. If you acquire the fluorescence images of fluorescent beads for Z direction and refer to it for deconvolution process, the images can improve moreover for reflecting optical properties of each microscope systems.
    Fluorescent beads is prefferable to PS-Speck Microscope Point Source Kit. It is enough to small - diamater is 0.17 micro meter - and contains four kinds of colors.

  • Before deconvolution, we recommend to prepare the original image for TIFF file type. Huygens can operate the original file type made by microscope or CCD camera, but it may not completely recognize them, especially multi dimensional acquisition (for example, multi channel time lapse).

  • It is ideal that original images are aquired at 16 bit depth. Images acquired 8 bit depth (range: from 0 to 255) is obviously too small SN retio to operate for image process, off course deconvolution.

Sample images by deconvolution

1.Wide field fluorescence images
2.Confocal images
3.3D rendering sample