Media 4 - Visualization of the preganglionic neuron dendritic field in the lumbosacral intermediotal lateral column

<p dir="ltr">Visualization of the<b> dendritic field</b> via retrograde filling of <b>lumbo</b><b>sacral (L6-S1)</b> preganglionic neurons in rat spinal cord. Microinjection of the retrograde tracer <b>cholera toxin subunit B (CTB)</b> into the <b>major pelvic ganglion (MPG)</b> of a female rat was taken up by sacral preganglionic neurons in the <b>intermediolateral column (IML)</b>. </p><p dir="ltr">Whole tissue immunolabelling and clearing using a modified form of <b>iDISCO </b>for spinal cord enabled the imaging of retrogradely-filled neurons. Large volume imaging was performed on the <b>light sheet microscope</b> with a 12x lens (Ultramicroscope II, Miltenyi Biotec). Punctate appearance of CTB granules along dendrites was segmented using the <i>Spots</i> function in Imaris (Bitplane), while the <i>Surfaces</i> function was used to segment the preganglionic neuron somata.</p>