The GolgiChromeTM kit (NDT107) is the first modified and commercialized version of the Golgi impregnation that allows simultaneous visualization of (a) the structural details, (b) the antigens’ characterization, (c) the anatomical interactions between discrete neuronal elements and (d) the 3D reconstruction and modeling of the nervous system through staining for simultaneous metal impregnation and immunohistochemistry.
NDT107 kit is suitable for 4% PFA perfused brains with the optimal preservation of antigenicity of the tissue, which can be co-localized with the iimmunoreactive proteins of interest and Golgi-impregnated neurons. Confocal laser scanning microscopy (CLSM) is recommended to use for acquiring to characterize the neurochemical as well as morphological properties of the individual neurons (Figure 1).
The following examples demonstrate the co-localizations of Golgi-cox impregnated neurons immunoreactive to Tyrosine Hydroxylase (TH), TH and Postsynaptic Density 95 (PSD-95) or Synapsin (SynI) of a fully impregnated pyramidal cell (Figure 2).
Figure 2.Golgi + TH + PSD95 or SynI immunohistochemistry of the prefrontalprelimbic zone of the rat brain, by Spiga et al (2012) (click to see enlarged photos)
Materials Required, but Not Included:
Double distilled or deionized water.
Plastic or glass tubes or vials.
Histological supplies and equipment, including gelatin-coated microscope slides, coverslips, staining jars, ethanol, xylene or xylene substitutes, resinous mounting medium (e.g. Permount®), and a light microscope.
Haug F-M S (1973) Heavy metals in the brain: a light microscope study of the rat with Timm’s sulphide silver method. Methodological considerations and cytological and regional staining patterns. Springer-VerlagBerlinHeidelbergNew York.
(Spigal et al 2011) Simultaneous Golgi-Cox and immunofluorescence using confocal microscopy. Brain Struct Funct. Sep 2011; 216(3): 171–182.