Monochrome conversion of colour immunofluorescence or in situ images

Here is the scenario. You have just spent ages constructing a beautiful multi panel figure with lettering and immuno or in situ micrographs. It all looks great the blue DAPI staining contrasts beautifully with the red target  staining. Here is a sub panel of one recently prepared by Teruhito.

And then, horror, you discover the journal will only publish the figures in monochrome. The default option is to export the figure as, say, a high resolution TIFF file (at least 300 dpi), open the image in photoshop (or the free, open source alternative GIMP, but I will use photoshop as the example here), and change the mode to grayscale. 

Here is the result. Alas, all those nice red highlights are grey, and look exactly like the converted blue bits. It is impossible to see which cells are stained positively and which are just bright blue DAPI staining. Blue and red are contrasting. Grey converted red is the same as grey converted blue.

But all is not lost. By using a more sophisticated approach to the conversion, you can mute the blue and bring up the red, as illustrated below.

Here is the same image with tweaked grayscale conversion. The DAPI staining is restrained allowing the red to show brightly.

I have attached a PDF file with basic instructions HERE. There are other methods that may work better, depending on your image and how you have created it. If you have used, say, Powerpoint to generate your layout, options for manipulating the colours within Photoshop are limited. Within Adobe Illustrator there are alternative approaches, as Illustrator has sophisticated colour management options. However learning enough about Illustrator to make a good layout and then manage colour within the application may take you several days or maybe weeks.  The approach I describe will take minutes.

Have fun.

attached file: Converting Immuno and in situ photomicrographs to monochrome.pdf