A10%solutionofFluoro-Rubyismadebydissolving10mgofFluoro-Rubydrypowderin100µLofdistilledwater.Itisthendeliveredviaintracranialstereotaxicinjectionusinga1uLHamiltonmicrosyringetopressureinjectthetracer.Injectionvolumestypicallyrangefrom0.02-0.1µLandaregraduallyinjectedovera10-15minuteinterval.Theanimalisallowedtorecoverandissacrificedtypically4-14dayslater.

Theanimalisthenperfusedwithneutralbufferedformaldehyde(10%formalinor4%paraformaldehydein0.1Mneutralphosphatebuffer.)Thebrainisremovedandpostfixedforatleastovernightinthesamefixativesolutionplus20%sucroseforcryo-protection.Thebrainisthencutonafreezingslidingmicrotomeorcryostatintosectionsusuallybetween20and40micronsinthickness.

Thetissuesectionsaremountedongelatincoatedslidesandareairdriedonaslidewarmerat50-60degreesCentigradeforatleastonehalfhour.ThedryslidesarethentransferredtoxyleneclearingsolutionforatleastoneminuteandthencoverslippedwithDPXmountingmedia.TheslidescanthenbeexaminedunderanepifluorescentmicroscopeusingafiltersuitableforvisualizingTRITC(greenlightexcitation).

Fluorochorome 公司于1985 年成立于美国科罗拉多州丹佛市，Fluorochorome 一直致力于荧光染料的研究与开发，其产品Fluoro Gold（荧光金）自 1985 年来在全球范围内被广泛应用，并有大量的参考文献。同时，Fluorochrome公司特别开发了高灵敏度的 Fluoro-Gold Antibody 、以及Fluoro-Ruby（红色荧光金）等产品，Fluorochrome 公司坚 持以优质的产品和热情的服务赢得了良好的知名度和口碑。