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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.
Under the new Analysis menu in Photoshop CS3 Extended is the new Place Scale Marker option. If the image is calibrated, this is a fast method to add a simple line scale marker to an image. If you are following along with me, open 40x_25u.tif and Myo40x.tif both found in your Chapter 11 Exercise files. The first thing we are going to do is custom calibration from our 40x_25 that we then can use to apply to the Myo40 and have Photoshop add the scale measurement marker. We go to Analysis > Set Measurement Scale > Custom.
We use the Ruler tool that it automatically gives us to draw across four of the grids, holding down our Shift key so it's horizontal, and we make sure we match the edge of the line to the right hand side of the grid line so it matches our beginning point. We see our Pixel length is 405, our Logical Length, we want to make sure it's 100 and then Logical Units. We want to change this so that it's Mu, and on the Macintosh, you hold down the Option key and hit M. Alternatively, you can hold down the Alt key and type 1254 on the number pad.
Now, that we have our micron symbol, the Mu. We can type a lowercase m and we are going to save this preset. Hit Save Preset and here we are going to type in 40x_100 then the Mu symbol, the lowercase m, and we hit OK. We can hit OK again, and go to Analysis and now, under our Set Measurement Scale, we have created an additional preset. Now, we can go to the Myo40x and select that. We can assign the correct scale by going to Set Measurement Scale and selecting the 40x_100 microns. We are going to let Photoshop place a scale marker for us. Go to Analysis > Place Scale Marker. In this case, it would give us a line of exactly 100 microns. If we wanted to put in 200-micron line, we would put into, for 500-micron line, we would add 5. So it just multiples the length by the actual scale that you have created. We can either have text automatically placed. We can position it either below or above the scale mark, and we can change the color to either black or white.
In this case, we will keep it black. We hit OK and Photoshop quickly runs through an action placing in the bottom left corner, the scale bar and the text that goes with it. We will select the Move tool by hitting V. Now, we can re-position this wherever it's needed in the image. Within the layer group is a layer for the marker graphic and a layer for the text, and we are able to modify each of these by selecting the specific layer.
And in this case, we have selected the marker graphic and we can double-click the layer thumbnail. We bring up the ability to change layer styles. In this case, we are going to use it to change the color of our graphic. We go to Color Overlay, select that, we double -click and now, we can select whatever color by hitting the color box. I would like to have this graphic yellow, because that's a very high contrast and it will show up very nicely on a screen. I add 255 of green to 255 of red to get to my pure yellow, I hit OK, and I hit OK again, and I have a yellow graphic line.
I also can change the text. I will click on the text box, and the Options bar gives me the ability to customize the text. In this case, 100 microns is too small to be legible. So I can increase its size, go to the Font size. I will approximately double it, and then I can select the Move tool and position the text to the middle and lastly, I will double-click on the text layer again and I will make sure I convert it to yellow simply by adding 255 of red and 255 of green. I hit OK. In that way, I have matched the colors and I have a legible text with my scale marker.
If I needed to, I could rotate the scale marker graphic by clicking on that layer, going to Edit > Transform, I can rotate it 90 degrees, and then I can move it along the side if that's what's needed. So you are able to use the automated feature of Photoshop, but then customize it as needed by working with the different layers.
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