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    • CommentAuthorjuruble
    • CommentTimeJan 31st 2008

    This protocol explains how to: add a scalebar to an image.

    Protocol revised: 1/30/2007

    Protocol written by: Barbara Lom

    1. To include a scale bar on any image you need to acquire the image and an image of the micrometer (tiny ruler) slide at the same magnification and image parameters. 
    2. Open your "ruler" image in Photoshop.  If the image is a bit askew you will need to rotate the image so that the ruler marks are "square" in the frame of your picture (it is OK to crop the ruler image if necessary in this step).
    3. Go to the "layers" box (usually at the lower right - if not then make layers visible under the "window" menu & select "show layers")
    4. Click on the arrow in the circle in the upper right corner of the layers box and select "new layer"
    5. Go to the toolbar and select the pencil tool (which is sometimes hiding under the paintbrush tool).
    6. Click on brush arrow (above the image) to change the "width" of the pencil lead (hint: select one of the "square" leads if possible - avoid the "circular" leads for a crisper scale bar – if you do select a round lead make sure it is a hard/all dark option and not one of the options that fades away at the edges). 
    7. Draw a line that corresponds to a reasonable and appropriate distance (which will depend on magnification).  Hint: hold down the shift key while you are drawing to get a perfectly flat line.  Note: your line will probably be black or white – whatever has the most contrast when placed on your image.  You can select a white or black line by clicking on the foreground options (top of the two overlapping squares on the toolbar)
    8. Now you will copy the line layer and move it to your growth cone image by going to the "select" menu and choosing the "all" command.  Then you will to the "edit" menu and select the "copy" command.
    9. Switch to your growth cone image.  Make a new layer and “paste” your scale bar over the growth cone.  You may need to use the move tool to get the scale bar into an appropriate location (usually the lower right corner).
    10. Use the text tool to add your distance – your text shouldn’t be any longer than the length of your scale bar line.  (the “m” in Symbol font will give you the μ for micrometers)
    • CommentAuthoraadeal
    • CommentTimeDec 14th 2010

    Note that the stereormicroscopes in the lab have different objectives. As indicated in step 1, it is important to the accuracy of your scale bar that you use the same stereomicroscope for your image and your micrometer image.

    • CommentAuthorsapyfrom
    • CommentTimeDec 15th 2010

    In case you're wondering where to find/return the micrometer, it goes on this cabinet:

    on this shelf!:

    • CommentAuthorcpgruber
    • CommentTimeSep 30th 2011
    Starting on step 9, position the scale bar in the bottom right corner of your image and in such a way that it is equidistant from the bottom and right sides of the image. If this placement interferes with a focal part of your image, move the scale bar either to the left or upward into a less important area of your image. You may also find that the color of your image makes it difficult to read both black and white scale bars. If this is the case, you can make your scale bar more apparent by adding a new layer behind the scale bar and its text, and using this layer to create a small solid white box (using the square shape tool) to give your scale bar greater contrast and readability.
    • CommentAuthormamccreery
    • CommentTimeSep 30th 2011
    When drawing your scale bar, begin and end at similar points on the micrometer bars (i.e. begin drawing the scale bar from the left side of one micrometer marker to the left side of the ending micrometer marker). Note that the size of your scale bar should be approximately one tenth of the length of the entire image.
    • CommentAuthoralfleming
    • CommentTimeOct 1st 2011
    When taking the picture of the micrometer make sure the image is straight across so when you draw the scale bar it is a straight line across and not diagonal. You can also rotate the line in photoshop to make sure it sure it is straight across in the image and not at an angle.