3.2.1 Triple-label immunocytochemistry
To confirm the presence of three proteins, TRPV1, EP4and B2, in same cell, triple-label immunostaining with EP4 (blue, 1:200), TRPV1 (green, 1:1000) and B2 (red, 1:200) were conducted. Confocal images shown in figure 6A illustrate immunolabelling for the three proteins of interest.
Morphometric analyses of six ganglia have shown that all the three proteins were present 21.7% of DRG neurons. The size-frequency histogram presented in figure 6B shows that triple-labelling was limited to small neurons (96.4%, <1,000 μm2), with only a small number of medium sized neurones labelled (3.6%, 1,000 -2,000μm2), and no large neurons (>2,000 μm2) labelled. The mean area of labelled neurons was 651.9 ± 15.2 μm2.