Metadata import problems
The Huygens Software reads different image File Formats. It will also try to extract from them the Microscopic Parameters necessary for Doing Deconvolution as much as possible.
The problem is that some file headers contain relevant information but not about the experimental conditions (which are the important ones for deconvolution) but about the microscope setup. It is up to the user to prepare the experiment in a way that is convenient for the acquisition settings.
For example, the microscope knows about its own filters, not about the WaveLength of the radiation coming from the fluophores. If the emission spectrum is centered in the filters or not is up to the user, but the microscope doesn't report it. Similarly with the excitation filters.
In the best case, looking at the file headers a final user can know about the wavelength range of the filters associated with each logical (data) channel, but the microscopes do not (always) report (properly) which laser line was used to excite the information acquired in that channel.
As a conclusion, the Huygens software makes a guess based on the available reported data: it assigns to each channel a value of the Emission Wavelength that is the center (average) value of a channel filter range, and find out an Excitation Wavelength that somehow fits together with that value.
Moreover the information is not always organized in the header the way we would like. Another difficulty is the channel order. Not always the logical order of the datasets match the reported laser lines order. That is why the previous matching becomes difficult sometimes.
It is always recommended to confirm that the guessing of the software corresponds to the real acquisition conditions.
