Fig. 3.

Whole mount RNAscope in situ hybridization (WH-FISH) can be combined with immunofluorescence and endogenous fluorescent protein such as tdTomato. The overview (a) and details (c, d) show that TrkC-tdTomato fluorescent protein can be combined with immunofluorescence using antibodies against tubulin. Note that the spiral ganglion neuron (SGN) cell bodies are nearly exclusively labeled with RNAscope (b) and that the RNAscope granules overlap with the tdTomato labeled spiral ganglion neurons cell bodies (SGN in a, d). Rendering RNAscope signal in such overlapping labeling into white dots (d) allows easier quantification. Note that anti-tubulin staining shows the radial fibers (RF) extending to the organ of Corti (OC) and also shows the intraganglionic spiral bundle (IGSB) at the borders of the spiral ganglion neurons. Abbreviations: IGSB intraganglionic spiral bundle, OC organ of Corti, RF radial fibers, SGN spiral ganglion neurons. Squares in a indicate approximate location of images c and d