MERGE method for fluorescence microscope photographs
We often see fluorescent photographs presented in articles as merge, as shown below. By merge, you can distinguish whether two (or more) fluorescent signals at the same location in a tissue or cell are overlapping or not. Today, let's talk about how to merge fluorescence microscope photos.
1,Basic principle
The basic principle is to recalculate the colour data of the same pixel in two (or more) equal-sized photos according to a formula to get a new colour. For example, the red and green of a fluorescent photograph overlap to form yellow. This algorithm is based on the RGB "add colour mode" (as shown below), and Photoshop's layer fusion mode "colour dropout" is almost the same.
2, software introduction
Today we introduce a scientific research in the common software: ImageJ, with it to merge photos than with Ps is more simple, more direct (do not care about layers, layer mode).ImageJ does not need to be installed, the software can even be opened on a USB stick to use.
There are two ways to merge fluorescence photos, one is to merge them with the software that comes with the microscope, and the other is to save the fluorescence photos of the same field of view with different excitation lights, and then merge them with the regular image processing software (e.g., PS) when presenting the results.If you are worried about quenching of fluorescence when looking at the fluorescence microscope (especially when stained by fluorescent dyes), you can take a bunch of photos quickly, and then choose the ones that are good when going back. When looking at the fluorescence microscope, if you are worried about fluorescence quenching (especially with fluorescent dye staining), you can quickly snap a bunch of photos first, and then go back and merge the ones that are good for the presentation of results. In this case, the second method may be more flexible and comfortable.
