// This macro demonstrates an example of Process>FFT>Custom Filter // command for removing horizontal interference streaks of different // amplitude in transmission mode confocal images. // Author: Gilles Carpentier, Faculte des Sciences et // Technologies, Universite Paris 12 Val de Marne // A sample image is available at // "/ij/macros/images/MyoblastCells.tif" // These are myoblast rat cells in early begining of differentiation, // in vitro. Acquisition by Gilles Carpentier. Dr Isabelle Barbosa // created the cell culture. // Image acquisition : // Confocal microscopy images of myoblasts cells were acquired // with a Zeiss LSM 410 laser scanning confocal Axiovert 135M // inverted microscopein in transmission mode with 488 nm // emission ray of a Ar/Kr used as lighting, no emission // filter was used. Debluring was performed before FFT treatment. //Comments: // Gilles Carpentier(1), Eric Delechelle(2) and Patrick Karasinski(2) // (1) Faculte des Sciences et Technologies, laboratoire CRRET, // CNRS, FRE-2412, Universite Paris 12-Val de Marne, // 61 Avenue du General de Gaulle, 94010 Creteil cedex, France. // (2) Faculte des Sciences et Technologies, laboratoire LERISS, // Universite Paris 12-Val de Marne, // 61 Avenue du General de Gaulle, 94010 Creteil cedex, France. //Methods were first developped with The NIH Image software. //Here is a description of the priciple of the method: // Removal of artifactual horizontal stripes was performed // using fast Fourrier transform (FFT): // First, a fast Harley transform procedure is performed on // a squared 512x512 image, with swapping the quadrants of // power spectrum. A filtering using a mask in the frequency // domain is performed [Bracewell,1984]. The vertical // rectangular area used as mask, is centred on the vertical // axis of symmetry, and covers about 1 % of the spectrum. This // mask, used as a filter (zero value), eliminates the periodic // structures on the Y axis whatever was their frequencies on // Y axis, and with a very low frequency in the X axis. // Furthermore, it centrered the histogram by subtracting // the mean grey level of the image. The inverse FFT was // performed after using a gaussian transition [Reeves, 1990]. // The final image was then free of horizontal periodic artefacts. // References : // Bracewell, R.N., The Fast Harley Transform, // Proc. IEEE. Vol. 72, /NÁ8, 1984. // Reeves A.A. Optimized Fast Hartley Transform for the // MC68000 with Applications in Image Processing. Master // of Science Thesis, Thayer School of Engineering. // Dartmouth College Hanover, New Hampshire, 1990. // delete any existing Transmission_Filter if (isOpen("Transmission_Filter")) { selectWindow("Transmission_Filter"); run("Close"); } // Runs faster in batch mode but this requires ImageJ 1.33o or later. requires("1.33o"); percent = getNumber("Percent of the half vertical axis of power spectra for hight frequencies:", 0.005); sdhight = getNumber("Standard Deviation of gaussian transition for hight frequencies:", 3.5); //setBatchMode(true); // to get the id number the active image to be treated: // see http://rsb.info.nih.gov/ij/developer/macro/functions.html imageid = getImageID(); //get the name and dimensions of the active image to be treated nomdimage = getTitle; imagey = getHeight(); imagex = getWidth(); // Create mask filter size =imagex; newImage("Transmission_Filter", "8-bit black", size, size, 1); //make a vertical rectangular (3x(size)) mask centered on the filter image width=3; halfwidth=(floor(width/2)); //position of the hight left rectangle selection //((size/2)-halfwidth); barre(((size/2)-halfwidth),0,width,size); // gaussian convolution matrix for a gaussian transition of filter matrice= matrix(9,40); selectWindow("Transmission_Filter"); it=4; for (nconv=0;nconv