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Veteran pharmaceutical research scientist and member of Adobe's Biomedical Image Advisory Group, Eric J. Wexler shares his experience creating detailed biomedical imaging in Photoshop CS3 Extended for Biomedical Research. Eric shows how to use Photoshop CS3's selection, analysis, and editing tools to evaluate an image's color composition, modify images for research, optimize exposure with levels and curves, transform images with layers, and compensate for acquisition problems and limitations. Eric also explains how to add reference information to images, annotate and optimize DICOM animations, and share finished images with colleagues. Exercise files accompany the tutorials.
NOTE: Actual biological research images are used for this title's examples. Some of these images, including those of internal organs and dissected animals, may be considered graphic or offensive to some viewers. Viewer discretion is strongly advised.
Often, we need to combine images side by side for comparison, control versus positive, a high resolution close up from a larger source image. By combining two images in a single reference, a reader is able to understand the direct comparison and appreciate the differences or lack thereof. If you are following along with me, open up in the Chapter 12 Exercise Files, ORO_Normal and ORO_Plaq. Here we want a comparison of two images, one a normal coronary vessel, the other one demonstrating atherosclerotic plaque. These are two relatively large images, both 5.5 megabytes, and we are going to reduce each of their sizes prior to combining them.
We'll first select the normal image. Go to Image > Image Size and we are going to reduce the Width to 3 centimeters and increase the Resolution at the same time to 300. In shrinking the image, we are reducing its size, but we are also increasing the Resolution slightly, so we'll still have an appropriate fineness of detail for this image. We hit OK and if we double-click on the magnifying glass, we can see this image at 100% by opening up the window.
We are going to do the exact same thing to the plaque that's been stained with Oil Right Out. Go to Image > Image Size, reduce the Width to 3 centimeters, at least double the Resolution to 300, hit OK. We can double-click on the magnifying glass and open that window up and we have our two images we are going to combine. They are both 3 centimeters in Width. By knowing that we can create a new canvas that we can fit both of these on as well as have appropriate spacing between the images.
We will go to File > New. The Resolution is 300, and we are going to make sure we select our Width in centimeters. In this case, we want to have a quarter centimeter between both images as well as on the other side. So we will make sure this is 7 centimeters and then the Height, we also want to make it large enough so that we are not cropping any of the image out. So in this case, if we typed 5, we are good and we will have a white Background.
We can give this a name, ORO Composite and then we hit OK. We would now turn on our Grid; it will help us align everything. And we can use the Move tool to simply drag the image down and over and we can do that for the plaque, we will grab that image, drag it down and over. And now between the Grids and the Smart Guide we can align this, so that they are in perfect relationship to one another. We do need to open up the Layers to be able to pick the separate layers to work with.
Click Layer 1 and we have these both aligned and we have our spacing equal on both the sides and between the two. So now we have our too images if we wanted to we can make them touch, we can put a black line between the two, but in this case we have this position the way we would like. We can get rid of some of the extra white space, by using the Crop tool and we crop away the area that we don't want. We can hit Return, and now we are going to refine this image and reduce its memory usage.
We can see on the bottom so that we can increase its efficiency to about 800 kilobytes from its current size 1.5 megabytes and that can be done by an easy flattening. Now we added the shortcut key, Ctrl+F or Command+F on the Macintosh and now we reduce its size and these are all on a single layer, we can save this as a JPEG, File > Save As, and we are going to go to select the Format to JPEG. We can save this as the largest JPEG file possible, hit OK.
Now we have an image that's ready for presentation on a PowerPoint slide or to share via email. Next, in this video using a single large high resolution image, we are going to create an insert, so that we can see a high resolution picture yet also know its location from a picture of the entire specimen. This is a plant stem and it started out as a 20 megabyte image that was created by a system that composites individual microscope images together. Now let's open up the image we are going to use to build this picture. If you are following along with me, open 20mbstem.tiff. To create the insert we are going to select a portion of this image and keep it at its original size and then we are going to reduce the size of the entire image and then replace that set-aside portion of the image back on to the same picture.
So we use our Square Marquee tool. Now in this case, I want to have a Fixed Ratio of a 4x6. So I type in the Width 6 and I type in Height 4. My ratio will always have at least those dimensions and I am going to copy this area. So I am going to do Ctrl+C or Command+C on the Macintosh. Then Ctrl or Command+N to create a new image, hit OK and then Ctrl or Command+V to place the original resolution fixture into it and on the side.
The next thing I need to do is mark that. This is the area that the image came from. So I will use the Stroke Command, go to Edit > Stroke. It doesn't need to be a very thick border, I will make this three, keep it black and I want this on the outside of the selection. I hit OK, now when I deselect, I have a square that exactly matches where I took the high resolution picture from. Next, I am going to resize this image, go to Image > Image Size, and I want to reduce its size to 10 centimeters and in this case, I can increase the Resolutions, I am not throwing away all of that data, while I resize it and that reduces the size of the image from 21 megabytes, down to three-and-a-half megabytes. I make sure that Bicubic Sharper is selected and that Constrain Proportions is on, so the image is not distorted, hit OK.
Now, I can take the image that I set aside, I can use the Move tool by hitting V and just drag it on top of the other image. In this case, I also want to have a border here. So I will go to Edit > Stroke and I add the same border to the outside, hit OK that strokes the image in Layer 1. The next thing I want to do is add an arrow, I go to my Save tools, pick my Align tool and I want to make sure that I confirm where the arrowhead is going to be placed.
In this case, we are going to put it at the end of the line and we want this to be a filled in arrow, so we make sure we have Fill pixels checked on. We might want to make this arrow a little bit more visible. We double the Weight of the line and now we are going to draw our line from the area that we took the image to the insert. The one last thing is we want to have a border around the entire image. So we select the Background, we double-click to change it to a normal layer, we hit OK.
Then we go to Edit > Stroke, and we can create a nice border. Increase the Thickness to 6 and make sure the location is inside. We hit OK and we have created a border around the entire image. Now we would want to save this as a Photoshop file, in case we want to make any changes later on, File > Save As. Go to Format and select Photoshop, modify its name, hit Save and we are able to go back any time and rework this image.
So that's two possible ways of combining two images. You can build upon this to add many images and create much larger and much more complex composites. In the next video, I will be demonstrating how we can use Bridge to show this work in an automated fashion like the built-in slideshow feature.
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