--- title: Intensity measurements layout: page --- ## Intensity measurements ### Activity: Measure intensities in image regions * Open image: xy_float__h2b_bg_corr.tif * Measure for both nuclei: * Maximum intensity * Average intensity * Median intensity * Sum intensity * Discuss the interpretation! * Discuss where to measure! ### Activity: Intensity measurements without pixel based background correction #### Motivation There are several good reasons not to subtract the background from each pixel in an image: * It is a bit tricky to do it right, because one has to convert to float to accomodate floting point and negative values. * If one has really big image data (TB) one would need (at least) another TB storage for the background corrected version of the image. #### Workflow * Open image: xy_calibrated_8bit__two_nuclei_high_background.tif * Measure for both nuclei and a background region: * Maximum intensity * Average intensity * Median intensity * Sum intensity * Discuss how to correct the intensities for the background * Appreciate that you also need the region areas for this task * Measure the region areas * Watch out: the image is calibrated! * Use the area for the correction. ### Formative assessment: Intensity measurements Fill in the blanks, using these words: integrated, mean, number_of_pixels, decrease, increase, sum 1. Average intensity is just another word for _____ intensity. 2. The _____ intensity is equal to the mean intensity times the _____ in the measured region. 3. In an 8-bit image, increasing the size of the measurement region can only _____ the sum intensity. 4. In a float image, increasing the size of the measurement region can _____ the sum intensity.