The merge method of fluorescence microscope photos
We often see in articles that fluorescent photos will be displayed in the form of merge, as shown in the figure below. Merge can be used to distinguish whether two (or more) fluorescent signals at the same location in a tissue or cell overlap. Today we will talk about how to merge fluorescence microscope photos~
1. Basic principles
The basic principle is to recalculate the color data of pixels at the same position on two (or more) sheets of equal size according to a formula to obtain a new color. For example, the red and green colors of a fluorescence photo overlap to make yellow. This algorithm is based on the RGB "addition mode" (as shown in the figure below), which is almost the same as Photoshop's layer fusion mode "screen color".
2. Software introduction
Today I will introduce a commonly used software in scientific research: ImageJ. Using it to merge photos is simpler and more direct than using PS (don't worry about layers, layer modes). ImageJ does not need to be installed, and the software can even be opened and used in a USB flash drive. For your convenience
There are generally two methods for merging fluorescent photos. One is to use the software that matches the microscope to merge, and the other is to save the fluorescent photos corresponding to different excitation lights in the same field of view. When displaying the results, use conventional image processing software ( For example, PS) to merge. If you are worried about the quenching of the fluorescence when looking at the fluorescence microscope (especially when stained with fluorescent dyes), you can quickly snap a bunch of photos, and then select the good photos to merge when you go back for the display of the results. At this time, the second method may be more flexible and calm.
