Author: Kavagiou Zaharoula (Kavagiouzaharoula@yahoo.com)
European Molecular Biology Laboratory, Heidelberg
Cell Biology and Cell Biophysics Programme
Light Microscopy GroupHistory: 2002/09/06: First version
Source: Patchwork_.java Installation: Download Patchwork_.java to the plugins folder, compile it with Plugins/Compile and Run, then restart ImageJ. Description: The program patchwork_ (running under ImageJ version 1.27z) is able to combine images that are overlapping (mosaic). The program was tested on bright transmission images coming from a microscope. With this program, the user can combine all the images of a mosaic into one. The program can handle 8bit, 16bit and color (RGB) images. The option of correcting the flaws of the images is also given (See Background Correction). The images should be aligned (which means that you don't have to shift them in 2 dimension to find the match, but only in one). A Few Guidelines The main program is the "patchwork_" one. All the other classes are used by this one. Class patchwork_ uses classes: ...correl2_ ...correl3_ ...Final_proc_ ...File Opener_ (downloaded from ImageJ PlugIn list) 1. Compile and Run patchwork_ .java 2. The user should provide the number of the images he/she wants to combine, in the beginning. 3. The user should open the images in the correct order (from the one in the right bottom to the one on the left top, moving upwards. Imagine the mosaic as a matrix of images and start from element (n,m), then open (n-1,m)...,(n,m-1),(n-1,m-1)...to (0,0). The File Opener plugin is used (as found on the webpage) in order to open the images. The user can also open the images before running the program and choosewhen the File Opener runs. 4. The user should provide the number of the "rows" of the mosaic, the percentage of overlap vertically and the percentage of overlap horizontally. 5. The option of eliminating background abnormalities is given. The images are divided by a "background image". Click OK to implement Background correction (for better results). If you don't want background correction, just change the default option and click OK. When the user opens the background image, the program starts combining the images. 6. Let it run until you are asked to save the final image or until an error message about the input appears. 7. Save the final image otherwise you lose it! Installation and Documentation In order to run the patchwork_ program to turn a mosaic into a single image, download the .java files of the classes involved (patchwork_.java, correl2_.java, correl3.java, Final_proc.java, File Opener.java). Compile each one of these files, ignoring the NullPointerException and canceling the File Opener_ run. Compile patchwork_ in the end and run it. The comments in these programs are written in such a way as to be easy to create a documentation file (HTML) by using Doxygen program (run the Doxygen wizard and use the above .java files as input). (download it from: http://www.stack.nl/~dimitri/doxygen/download.html#latestsrc) Detailed explanation of the purpose of patchwork_ program When we want to examine an object (e.g. cell) with a microscope, we prefer to zoom in in certain areas of this object to get better resolution, but we also want to have the whole picture. So we create a mosaic by capturing parts of the object and getting images that overlap (we need the overlap to reconstruct the object again). Reconstructing the whole image from some smaller ones is the purpose of this program. Background Correction When capturing bright transmission images the result is not always satisfying. For example, there may be stains on the lenses and, usually, the lighting is not homogenous all over the image. These may cause problems when we try to reconstruct the whole picture. In this program, there been an effort to correct this by taking an image with no object inside (with the microscope), which unavoidably has all the flaws described above. We divide this image by every other image that is part of the mosaic and we get corrected images to combine. Example If you want to view a little demonstration of this program, you can use the images that are in the following address. www.photos.yahoo.com/kavagiouzaharoula (Image folder) Download them and save them in the order they are numbered so that you can open them easily when you run the program. The mosaic is created by 4 8bit grayscale images and you can use the "background image" to correct the other 4. The correct answers for the program are: number of images -> 4 number of rows -> 2 vertical overlap -> 40 (%) horizontal overlap -> 27 (%)