This Kit I is designed for the detection of degenerating neurons in fixed tissue sections of the central nervous system of experimental animals. The principle of this kit is based on the findings that some components of neurons that undergo degeneration, such as lysosomes, axons, and terminals, become particularly argyrophilic. Under certain conditions, these cellular elements bind to silver ions with high affinity. After reduction, the silver ions from the metal grains are visible under an optical or electron microscope.
The NDT103 kit has been widely used in animal studies under various experimental conditions1-16. This kit has proven to be extremely specific and sensitive for the detection of degenerating cells, axons, and neuronal terminals in both the brain and spinal cord (see photo below). It is particularly useful for the detection of small numbers of degenerating neurons that may not be demonstrable with routine histopathological techniques.
Neurodegeneration detection with NeuroSilver Kit (NDT103).
A: section (40 microns) through the dentate gyrus of the hippocampus of a rat injected with kainic acid (10 mg/kg, sc), showing neurons and degenerate processes (black) in the polymorphic layer (pl) and the layer molecular (ml), respectively.
B: coronal section (30 microns) through the septum of a rat, sacrificed 10 days after a unilateral section of the fimbria. Note the degenerated axons (indicated by the arrow) in the ipsilateral cul-de-sac.
C: horizontal section (40 microns) through the dentate gyrus of the hippocampus of a dead rat at 5 days after intraentorhinal injection with aminooxy acetic acid. Note the numerous terminals of degenerating axons (arrowheads) in the middle of the molecular layer (ml), the terminal field of entorhinal neurons killed by drug injection (for more details see Neuroscience 82: 1165, 1998).
D: coronal section (40 microns) through the lateral geniculate nucleus of a dead rat at 5 days after a unilateral aspiration of the visual cortex. Observe the terminals of dense degenerate axons in the subgeniculate nucleus (SubG) (Section courtesy of Dr. E.-Y. Chen, Rush University [D])
Detection of neurodegeneration in the rat brain from a stroke model.
A 40-micron cryostat section was coronally cut through the striatum of a rat used as an animal model of stroke. The section was processed to demonstrate neuronal damage with the NDT103 kit. Note the accumulation of metallic silver grains (black) in both the striatum and the cortex, indicating the presence of neurodegeneration.
Detection of neurodegeneration in the rat brain from an epilepsy model.
40-micron cryostat section through the cortex of a rat used as an animal model of epilepsy. Note the degenerating (black) neurons in the deep layers.
Detection of neurodegeneration and amyloid plaques in the mouse brain from a model for Alzheimer’s disease.
A 40-micron cryostat section was cut coronally through the dorsal hippocampus of a transgenic mouse used as a model to study Alzheimer’s disease. The section was processed to detect both neuronal damage and amyloid plaques with the NDT 103 kit. Note the silver-stained plaques in both the hippocampus and the cortex.
Detection of neurodegeneration in the rat brain from a model for Huntington’s disease.
A 40-micron cryostat section was coronally cut through the striatum of a rat that received a focal injection of quinolinic acid. The section was processed first to determine bcl2 immunoreactivity and then to detect neuronal damage with the NDT103 kit. Note that degenerating neurons (black) intermingle with surviving bcl2-positive neurons.
- Solution A (500 ml)
- Solution B (500 ml)
- Solution C (500 ml)
- Solution D (500 ml)
- Solution E (2 ml)
- Solution F (3 ml)
- Solution G (5X) (500 ml)
- Glass Sample Retriever (2)
Materials required, but not included:
- Bidistilled water
- Tissue culture plates (6 wells)
- Histological equipment and supplies, including gelatin-coated microscope slides, glass coverslips, hairbrush, staining bottles, xylene or xylene substitutes, and a light microscope.