Quantification High Pass Method Photoshop ImageJ

This video shows how to prepare an image for subsequent image analysis using Photoshop and Image J (FIJI). Quantitation steps include the Sedgewick method for segmenting features of interest from surrounding tissue for scientific images. This method allows for a consistent threshold value for images that are at different contrasts and brightnesses. Quantization (or quantify) includes the use of the bandpass (ImageJ) or high pass (Photoshop) filter to effectively segment smaller features at various sizes.

You can learn about classifiers when using Image J to eliminate thresholded features by using the circularity and pixel area settings. Image measurements can then be refined so that non-specific features are eliminated.

Get lengths via skeletonize, areas, and other measurements such as caliper length, bounding box, perimeter, etc. Use the make binary, skeleton analysis, ROI (Region of Interest), Minimum, Maximum, Median and other tools for quantification.

This is an in-depth learning video with three demonstration images. You will likely not come across these techniques in classrooms. Few instructors emphasize the importance of uneven illumination when segmenting according to tones in science images, such as those from confocal, electron and compound microscopes.