Quantification of Dorsal Root Ganglion Neurite Growth in a Multi-Compartment (MC) Device Using Fiji

After imaging the dorsal root ganglion cultured in a multi-compartment device, launch Image J and open the image file. To enhance the contrast and make the smallest neurites visible using the shortcuts Control with Shift and C, open the Brightness and Contrast option. Adjust the brightness and contrast sliders to visualize the neurites.

Then click on Apply. Now, navigate to Plugins, select Segmentation, and click Simple Neurite Tracer to open the neurite tracer. Then, click on one end of the neurite at the dorsal root ganglion body to initiate a new trace, and continue adding points along the neurite until it is fully traced.

Then click on Complete Path. Using the straight line tool in Fiji, draw a line of the same length as the scale bar without releasing the end of the line. And note the length value under the toolbar.

Set the scale to convert the neurite length to real units and copy the results to Excel. Then, convert the lengths to real units in Excel. Then, measure the length of six long neurites extending from both sides of the dorsal root ganglion, and calculate the average length and standard deviation of the dorsal root ganglion.

On days 27 and 21, there was robust growth of neurites in the multi compartment and plain gels, respectively. The average length of neurites in the multi compartment device was comparable to neurite length in control plane gels.