Axon Growth and Regeneration by Andrew J. Murray

Author:Andrew J. Murray
Language: eng
Format: epub
Publisher: Springer New York, New York, NY

8.Parameters such as number of neurites, neurite length, branching, and orientation are commonly features of interest. Immunostained neurites can be identified as objects using automated image analysis software, or in some cases it can be beneficial to trace neurites of interest manually (although it is important to note that if this is done using image projections or extended focus views then features in the z-dimension may not be apparent). A potential limitation with the use of automated thresholding to select neurites is that minor discontinuities in staining can result in single neurites being wrongly identified as multiple separate objects. Some image analysis software will allow similar adjacent objects to be merged automatically, but it is useful to check images carefully to ensure neurites are selected in their entirety. Where the direction of neurite growth is of interest, ensure that the image orientation is appropriate to allow a “bearing” or angle of deviation from one axis of the image to be relevant (Fig. 2b). Where absolute neurite length is required then images must include the cell body and entire neurite, which requires sampling protocols to take this into account.

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