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    • CommentAuthorjakhan66
    • CommentTimeMay 16th 2013 edited
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    Written by: Jannat Khan

    Revised on: 5/16/2013

    This method was used to measure the neurite length of PC12 cells after treatment with neuronal growth factor (NGF). However, the method may be utilized to account for the length of any morphological feature.

    Setting the Scale

    1.            Click File > Open and open an image of a micrometer.

    2.            Draw a line using the straight line tool between the known increments on the micrometer.



    3.            Click Analyze > Set Scale. Enter the length of the scale bar as Known distance and um as Unit of length. Click OK and the image size will be calibrated.

    ***Note: To calibrate all of the images opened in the current session, check the Global box before clicking OK.



    Tracing neurites

    1.            Click File > Open and open image of neurites that you wish you measure. 

    2.            Select the Freehand tool to trace a neurite (Neuron of interest boxed in black in the image).

    Hint: To select the Straight line button and right click to change to Freehand line


    3.            Using the mouse, move the arrow to the beginning of the neurite (sprouting from the soma). Hold left-click using the mouse and guide the arrow to follow the shape of the neurite. At the end of the neurite release left-click on the mouse.

    4.            Press COMMAND + M (Mac) or CONTROL + M (PC) to measure the traced neurite. Another window will appear with the measurement (Red circle = Length). As you proceed to count all the neurites in the pictures and the measurements will be accumulated in the window.