Diffusion and imaging properties of three new lipophilic tracers, NeuroVue™ Maroon, NeuroVue™ Red and NeuroVue™ Green and their use for double and triple labeling of neuronal profile

We describe here diffusion and imaging properties of three new lipophilic tracers, NeuroVue™ Maroon (near infrared), NeuroVue™ Red and NeuroVue™ Green. Using pair-wise comparisons between the new dyes and existing dyes (DiI, DiA, DiD, DiO, PKH2, PKH26) applied to the left and the right side of fixed spinal cord preparations, we show that NeuroVue™ Maroon (excitation maximum 647 nm) surpasses all other dyes in this study in signal to noise ratio. We also present data showing the utility of these new dyes for both double labeling and triple labeling in combination with each other or existing lipophilic tracers. Using mice bearing the PLP–eGFP transgene, we demonstrate that either NeuroVue™ Maroon or NeuroVue™ Red can readily be combined with eGFP labeling. Double labeling experiments using NeuroVue™ Red and eGFP allowed us to demonstrate that every fiber in the neonatal ear is surrounded by developing Schwann cells.