NeuroVue® Dye Filter is useful for neuronal tract tracing studies of up to 3-4 weeks. Spectrally compatible with eGFP and YFP in many systems, and NeuroVue Maroon, & Jade. Spectral unmixing required for use with NeuroVue Red.
Advantages:
Convenient, ready-to-use coated filter format
More precise control of dye insertion point
No messy oils, pastes or hard-to-position crystals
Diffusion properties comparable to or better than other commercially available neurotracing dyes
More focal results (e.g. labeling of small sets of axons within a pathway)
Available in multiple colors, including far red, for multi-tract tracing and improved results even in tissues with high myelin expression
Applications:
NeuroVue® Orange has been found to be useful for tracing neuronal connections in animal tissues fixed in formaldehyde (personal communication, Dr Bernd Fritzsch, Creighton University). Like other lipophilic tracers (1, 2), it readily transfers into plasma membranes in fixed and/or live tissues and diffuses laterally within the membrane, eventually labeling the entire cell body as well as the finest axonal and dendritic branches, and allowing visualization of neuronal processes up to several millimeters distant from the point of dye insertion. Studies of up to 3-4 weeks can be performed in most cases without substantial transcellular diffusion.
NeuroVue® Orange is provided in coated filter format because insertion of small dye coated filter segments has been shown to be a simple, reliable method for labeling well defined tissue regions, avoiding known artifacts associated with labeling via high pressure microinjection or insertion of dye crystals on a dissecting needle (1, 3, 4). NeuroVue Orange fluoresces in the red and exhibits minimal bleed through into filter windows typically used for far red fluorescing lipophilic tracers such as NeuroVue Maroon and NeuroVue Burgundy and green fluorescing lipophilic tracers such as NeuroVue Jade making it an excellent choice for multi-color neural tracing studies in sections and/or whole mount preparations (4). NeuroVue Orange can also be used in combination with NeuroVue Red using spectral unmixing techniques.
Additional Information:
1. Filter segments of the desired size and shape can be cut using super fine Vannas scissors and inserted into the tissue at the site to be labeled.
2. Diffusion times vary depending on the biological system under study and must be determined empirically. See Technical Data Sheet for references and protocol for potentially important variables and possible starting conditions.
3.Detection of Labeled Cells
Confocal microscopy: Detection is most efficient using the 543nm laser line for excitation and emission filter set at 565-615nm.
Epifluorescence microscopy: Standard filter sets potentially useful for NeuroVue Orange excitation and emission include:
Chroma 31002 : TRITC (Rhodamine)/DiI/Cy3 ®. Exciter D540, Dichroic 565DCLP, Emitter D605/55m.
Chroma 41002 : TRITC (Rhodamine)/DiI/Cy3 ®. Exciter HQ535/50x, Dichroic Q565LP, Emitter HQ610/75m.
Description:
1cm² filter coated with the lipophilic reddish orange emitting dye, NeuroVue® Orange (ex max=550nm; em max=570nm).
References:
1. Honig M. 1993 DiI Labelling. Neuroscience Protocols 93-050-16-01-20 .
2. Köbbert C, Apps R, Bechmann I, Lanciego JL, Mey J,Thanos S. 2000. Current concepts of neuroanatomical tracing. Progress in Neurobiology 62: 327-351.
3. Fritzsch, B, Nichols DH, Echelard Y, McMahon AP. 1995. Development of midbrain and anterior hindbrain ocular motoneurons in normal and Wnt-1 knockout mice, J Neurobiol. 27:457-469.
4. Rosa-Molinar E, Proskocil BJ, Ettel M and Fritzsch B. 1999. Whole-mount procedures for simultaneous visualization of nerves, neurons, cartilage and bone. Brain Res. Protoc. 4, 115-123.
Recent Publications:
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. B Fritzsch et al. Brain Res Bull., 66, 3, 249-258 (2005).
A disorganized innervation of the inner ear persists in the absence of ErbB2. JK Morris et al. Brain Res.,1091:186-199 (2006).
Deafferentiation induces novel projections in the auditory brainstem after hearing onset. CY Hsieh and KS Cramer. Journal of Comparative Neurology, 497:589-599 (2006)
Al-Mehdi AB, Patel M, Haroon A, Reed D, Ohlsson-Wilhelm B, Muirhead K and Gray BD. Increased Depth of Cellular Imaging in the Intact Lung using Far Red and Near Infrared Fluorescent Probes. Int. J. Biomed Imaging. 1-7 (2006).
Stewart CC, Woodring ML, Podniesinski E and Gray BD. Flow cytometer in the infrared: inexpensive modifications to a commercial instrument. Cytometry, Part A, 67A, #2, 104-111, 2005
Sold under sublicense from PTI Research, Inc. to MTTI . NeuroVue® is a trademark of PTI Research, Inc.