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Concept
Fluoro-jade stain
Summary
Fluoro-jade stain is a fluorochrome derived from fluorescein, and is commonly used in neuroscience disciplines to label degenerating neurons in ex vivo tissue of the central nervous system. The first fluoro-jade derivative was reported by Larry Schmued in 1997 as an alternative method from traditional methods for labeling degenerating neurons such as silver nitrate staining, H&E stain, or Nissl stain. Fluoro-jade may be preferred to other degenerative stains due to simplicity of staining procedures and visual interpretation, which are common drawbacks of conventional degenerative stains. However, the mechanism by which fluoro-jade labels degenerating neurons is unknown thus creating some controversy to the actual physiological condition of the labeled cells.
Currently, there are three fluoro-jade dyes (fluoro-jade, fluoro-jade B, and fluoro-jade C ), all of which are anionic derivatives of fluorescein and highly acidic. Specifically, fluoro-jade is a mixture of 5-carboxyfluorescein and 6-carboxyfluorescein disodium salts, whereas fluoro-jade B is a mixture of (1) trisodium 5-(6-hydroxy-3-oxo-3H-xanthen-9yl)benzene, 1,2,4 tricarboxylic acid, (2) disodium 2-(6-hydroxy-3-oxo-3H-xanthen-9yl)-5-(2,4-dihydroxybenzol)terepthalic acid, and (3) disodium 2,5-bis(6-hydroxy-3-oxo-3H-xanthen-9yl)terepthalic acid. All three fluoro-jade species have similar excitation and emission profiles as fluorescein (excitation: 495 nm; emission:521 nm) and thus can be visualized using a fluorescein/FITC filter. The newer dyes, fluoro-jade B and fluoro-jade C, were developed to improve signal to noise ratio, therefore creating superior compounds for visualizing finer neuronal morphology including dendrites, axons and nerve terminals.
Nearly all processed tissue is compatible with fluoro-jade stain including tissue from rodents (mice and rats), non-human primates and humans. Mounted tissue is stepwise rehydrated with decreasing concentrations of alcohol. Potassium permanganate may be used to decrease background staining and protect tissue from fading and photo bleaching.
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