The aligned data is generated as well, if desired. The alignment compensates for translation and magnification differences between the different wavelengths. Currently, unprocessed data do not compensate for magnification and translation diffences between the different wavelengths used in data collection. This can result in problems analyzing data when it is important to know how differently-labeled objects are positioned with respect to each other. This program uses a simplex to minimize the r-factor calculated from differences between these images.
All data is loaded into memory as two byte integers to reduce data storage sizes.
Basic usage Record dataset with either additional reference bleedthrough images or reference beads located within the dataset. The dataset can be spread over two files if more than 5 wavelengths are recorded. If there is more than one file, the -in2 option must be used. Process data through ccdcor and decon, as with any other dataset. Select the reference wavelength, align wavelengths, and compare wavelengths.