What are the characteristics of the Confocal, Widefield, Multiphoton (and other) modules?
How does each module work and what are is advantage in using these modules for each type of image.
To start: the time, multi photon options are independant, i.e. they have no overlapping features.
Multi-photon
There are 'widefield' and 'confocal' multi photon microscopes; <br>
the multi photon option gives access to both. <br>
Two (or multi) photon data has more in common with confocal data than with <br>
wide field data, but still is handled differently. <br>
For instance: <br>
- different PSF generator, though it shares parts with the confocal generator. <br>
- the reconstruct-PSF toold uses different a-priori knowledge about what the PSF should like <br>
- a number of basic values is computed differently, for instance the Nyquist rate, the expected two-point resolution, and so on. <br>
As a result all tools wich have something to do with optical properties behave differently. For example, average beads <br>
uses different criteria to reject or accumulate beads, background searches are conducted differently. <br>
- The restoration algorithms are in broad lines the same but differ in the way artifacts are suppressed and image edges are handled.
Confocal/widefield <br>
In the same way as for the multi photon case, all tools related to optical properties behave different. <br>
For instance it is possible to estimate the numerical aperture from wide field bead images whereas in confocal or two-photon it is not. <br>
Wide field data is in all restoration tools inspected for bleaching since not correcting it leads to poor results; the same holds for corrections of lamp instability of the lamp. <br>
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Time <br>
Processing time series is not simply a matter of frame by frame deconvolution: <br>
-the time-dependent background is determined and corrected for <br>
-for all types of data an attempt is made to find a bleaching curve over time. <br>
If this is found in the confocal or two-photon case it is corrected both over time and z. If it fails <br>
the bleaching is corrected over time, not z. The same applies for <br>
wide field, but there the bleaching is always corrected along z. <br>
-lastly is the deconvolution of the series <br>
itself, if necessary split into bricks, and for all channels.
In addition, to process time series with the 4D gaussian, laplacian, plotFLux, backbgound, 4D fft you need the time option.
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Keywords: option modules<br>
Categories: Faq Deconvolution, Faq Microscopy, Huygens Faq, Imported Faqs<br>
Platforms: Irix Linux Windows Mac AIX<br>
Related products: Hu Ess Hu Pro Hu Script<br>
