Cochlear neuronal tracing for frequency mapping with DiI, NeuroVue, and Golgi methods.
ABSTRACT Labeling experiments using NeuroVue Red dye allowed us to demonstrate individual afferent fiber tracks in the cochlea from the synaptic region of the inner hair cell in the organ of Corti (OC) to the spiral ganglion in Rosenthal's canal. Further optimization is necessary to obtain 3-dimensional (3D) neural distribution in the apical region for frequency mapping.
We intend to develop a method by which the radial fibers of the spiral ganglion (SG) can be individually visualized and tracked in 3D from the base to the apex of the cochlea. The combined trajectories of fibers from each cochlea could then be calculated for modeling of the 3D relationship of OC and SG in cochlear implant studies to assist in the optimization of cochlear implants for music and speech perception in noise.
We tested three different methods to visualize cochlear nerve fibers from OC to SG. Adult rat and mouse ears were stained with DiI dye, modified Golgi-Cox method or NeuroVue dye, sectioned or whole-mounted, and viewed with confocal or standard light microscope.
In DiI staining, spacial resolution and the number of neurons to be stained are too low to utilize this method to create a characteristic frequency map of the cochlea. The Golgi method mainly stained efferent nerve fibers, resulting in less information on cochlear nerve distribution. NeuroVue Red dye allowed clear tracking of individual fibers when combined with DAPI counterstaining.
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ABSTRACT: Adoptive cellular immunotherapy (ACI) has been demonstrated to be a promising cancer therapeutic, however, the distribution of immune cells injected into a tumor-bearing body is unclear. In this study, we investigated the tumor-targeting capacity of cytokine-induced killer (CIK) cells and cytotoxic T lymphocytes (CTLs) in a human gastric carcinoma orthotopic mouse model using a near-infrared fluorescence imaging system. CIK cells and tumor-specific CTLs were prepared with the near-infrared fluorescent dye DiR. As expected, no significant change in the proliferation rate or antitumor activity of CIK cells and CTLs was noted after labeling with DiR. Furthermore, a gastric carcinoma orthotopic model was established using a fibrinogen-thrombin method in nude mice followed by intraperitoneal infusion of the labeled immune cells into nude mice with established gastric carcinoma. Dynamic tracing of the immune cells was performed using a fluorescence-based live imaging system. Concentrated fluorescence signals were observed for a minimum of two weeks at the tumor site in mice infused with either CIK cells or CTLs with a peak signal at 48 h. Notably, CTLs were more persistent at the tumor site and exhibited a more intense antitumor activity than CIK cells following infusion. These results provided visual evidence of the tumor-targeting capacity of immune cells in live animals.Experimental and therapeutic medicine 08/2012; 4(2):221-225. · 0.94 Impact Factor