Useful for tract tracing studies of up to 3-4 weeks. Spectrally compatible with eGFP, YFP in many systems, and NeuroVue® Maroon and Jade. Spectral unmixing required for use with NeuroVue® Orange. NeuroVue® Red Plus provides faster and more efficient neuron labeling with higher dye loading than standard NeuroVue® Red filter square.
Product Description: 1 cm2 nylon filter coated with the lipophilic red emitting dye, NeuroVue Red.
Typical dye loading: 18-21nmoles/mm2.
Spectra of NeuroVue Red (ex max=567nm; em max=588nm)
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
- NeuroVue® Red Plus provides bright, clean, crisp neuron labeling with higher dye loading than standard NeuroVue® Red filter square.
Applications:
The NeuroVue Red Plus filter has a higher dye loading than the standard NeuroVue Red filter and can provide more extensive, bright, clean, and crisp neuron labeling with shorter diffusion times (1). NeuroVue Red has been found to be useful for tracing neuronal connections in animal tissues fixed in formaldehyde (2, 4-7, 9, 10, 12, 14, 15). Like other lipophilic tracers (8, 11), 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 (2, 4-7, 9, 10, 12, 14, 15). NeuroVue Red 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 (3, 8, 13). NeuroVue Red fluoresces in the red (Figure 1) and exhibits minimal bleed through into filter windows typically used for green fluorescing lipophilic tracers such as NeuroVue Jade and far red fluorescing lipophilic tracers such as NeuroVue Maroon or NeuroVue Burgundy, making it an excellent choice for multi-color neural tracing studies in sections and/or whole mount preparations (2, 4-7, 9, 10, 12, 14, 15). In addition, NeuroVue Red can be used in combination with NeuroVue Orange if spectral unmixing techniques are employed.
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.
3) Detection of Labeled Cells
Confocal Microscopy:
Detection is most efficient using the 543nm or 568nm laser line for excitation and emission filter set at 565-615nm.
Epifluorescence Microscopy:
Standard filter sets potentially useful for NeuroVue Red excitation and emission include:
• Chroma 41034 : Rhodamine X (or Alexa Fluor 568T) Exciter HQ570/20x , Dichroic Q585LP, Emitter HQ620/60m
• Chroma 31002 : TRITC (Rhodamine)/DiI/Cy3®, Exciter D540/25x , Dichroic 565DCLP, Emitter D605/55m
• Chroma 41002 : TRITC (Rhodamine)/DiI, Exciter HQ535/50x , Dichroic Q565LP, Emitter HQ610/75m
References:
1. Personal communication, Karina Cramer laboratory, University of California, Irvine.
2. de Caprona MD, Beisel KW, Nichols DH, Fritzsch B. 2004. Partial behavioral compensation is
revealed in balance tasked mutant mice lacking otoconia. Brain Res Bull 64:289-301. Both NeuroVue
Maroon (previously PTIR271) and NeuroVue Red (previously PTIR278) were used in Figure 8 (B.
Fritzsch, personal communication).
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. Fritzsch B, Muirhead KA, Feng F, Gray BD, Ohlsson-Wilhelm BM. 2005. 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. Brain Res Bull 66:249-258. NeuroVue
Maroon, NeuroVue Red, NeuroVue Green
5. Fritzsch B, Matei VA, Nichols DH, Bermingham N, Jones K, Beisel KW, Wang VY. 2005. Atoh1 null
mutants show directed afferent fiber growth to undifferentiated ear sensory epithelia followed by
incomplete fiber retention. Dev Dyn, 233: 570-583. NeuroVue Maroon (previously PTIR271),
NeuroVue Red (previously PTIR278)
6. Fritzsch B, Jackson Lab Presentation, 2005:
http://www.biomedsci.creighton.edu/facilities/nccb/media/Jackson_lab_presentation.ppt
NeuroVue Green (previously PTIR281);NeuroVue Red (previously PTIR278); NeuroVue Maroon
(previously PTIR271)
7. Gurung B, Fritzsch B. 2004. Time course of embryonic midbrain and thalamic auditory connection
development in mice as revealed by carbocyanine dye tracing. J Comp Neurol 479:309-327. NeuroVue
Maroon (previously PTIR271), NeuroVue Red (previously PTIR278)
8. Honig M. DiI Labelling. 1993. Neuroscience Protocols 93-050-16-01-20
9. Hsieh CY, Cramer KS. 2006. Deafferentation Induces Novel Axonal Projections in the Auditory
Brainstem After Hearing Onset. J Comp Neurol 497: 589-599 NeuroVue Red was used for all figures
except Figure 2D, for which both NeuroVue Red and DiI were used, and Figure 5A, for which DiI was
used (K. Cramer, personal communication).
10. Hsieh CY, Hong CT, Cramer KS. 2007. Deletion of EphA4 Enhances Deafferentiation-Induced
Ipsilateral Sprouting in Auditory Brainstem Projections. J Comp Neurol 504: 508-518.
11. Köbbert C, Apps R, Bechmann I, Lanciego JL, Mey J, Thanos S. 2000. Current concepts of
neuroanatomical tracing. Progress in Neurobiology 62: 327-351.
12. Morris JK, Maklad A, Hansen LA, Feng F, Sorensen C, Lee KF, Macklin WB, Fritzsch B. 2006. A
disorganized innervation of the inner ear persists in the absence of ErbB2. Brain Res. 1091: 186-199
NeuroVue Maroon, NeuroVue Red
13. 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 .
14. Tessarollo L, Coppola V, Fritzsch B. 2004. NTF3 replacement with brain-derived neurotrophic
factor redirects vestibular nerve fibers to the cochlea. J Neurosci 24:2575-2584. NeuroVue Maroon
(previously PTIR271), NeuroVue Red (previously PTIR278)
15. Zou D, Silvius D, Fritzsch B, Xu PX. 2004. Eya1 and Six1 are essential for early steps of sensory
neurogenesis in mammalian cranial placodes. Development 131:5561-5572. NeuroVue Maroon (previously PTIR271) and NeuroVue Red (previously PTIR278) were used for Figure 6, panels G-R (B. Fritzsch, personal communication).
Sold under sublicense from PTI Research, Inc. to MTTI . NeuroVue® is a trademark of PTI Research, Inc.