FluoMergingTutorial copy

FluoMergingTutorial copy - Merging uorescence images Hernan...

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Merging fluorescence images Hernan G. Garcia March 30, 2009 1 Introduction In this tutorial we will show how to merge the images of a fluorescent sample taken using different channels into one composite color image. When quan- tifying images it is usually better to work with the independent channels. However, it useful to be able to display all channels at once. Combining dif- ferent channels can also be useful when looking for colocalization of different floruophores. 2 Loading and setting up the images We start by loading the images corresponding to the three fluorescent chan- nels. You’ll be most likely working with a DAPI, FITC, and TRITC snaphots of the same cell. We’ll assign DAPI to the blue channel, FITC to the green channel, and TRITC to the red channel. This assignment is related to the actual wavelengths of the emission of the fluorophores. ImR=imread(’brain_60x_700ms_TRITC_wheels.tif’); ImG=imread(’brain_60x_400ms_FITC_wheels.tif’); ImB=imread(’brain_60x_200ms_DAPI_wheels.tif’); Remember that these images have a bit depth of 16 bits and that the range
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