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Fluo-8, AM 鈣離子熒光探針CalciFluor™ Fluo-8, AM 的應(yīng)用
CalciFluor™ Fluo-8, AM 已開發(fā)用于改善細(xì)胞負(fù)載和鈣響應(yīng)，同時保持便利的 Fluo-3 和 Fluo-4 光譜波長，最大激發(fā) @ ~490 nm 和最大發(fā)射 @ ~520 nm。 與 Fluo 4 AM 類似，陰離子羧基被乙酰氧基甲酯保護(hù)，允許膜滲透，隨后被細(xì)胞內(nèi)酯酶去除 - 允許 Ca2+ 檢測。

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CalciFluor™ Fluo-8, AM has been developed to improve cell loading and calcium response while maintaining the convenient Fluo-3 and Fluo-4 spectral wavelength of maximum excitation @ ~490 nm and maximum emission @ ~520 nm. Similarly to Fluo 4 AM, the anionic carboxy groups are protected with acetoxy methyl esters allowing membrane permeability, which are subsequently removed by intracellular esterases - allowing Ca2+ detection. However CalciFluor™ Fluo-8, AM is efficiently loaded into cells at room temperature (whereas Fluo-3 and Fluo-4 require much more harsh loading conditions), and CalciFluor™ Fluo-8, AM is 2 times brighter than Fluo-4 and 4 times brighter than Fluo-3. These features, as well as a lower price, make CalciFluor™ Fluo-8, AM a superior in vivo calcium indicator.

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FLUO-8 (AM) 是 Fluo-8 的一種可滲透細(xì)胞的乙酰氧基甲基 (AM) 酯衍生物。 亮度大約是 Fluo-4 的兩倍。 吸光度 452 nm (26,000)； 發(fā)射 515 nm（AM 水解后；溶于 10 mM EGTA、100 mM KCl、10 mM MOPS，pH 7.2）

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Fluo-8 AM, green fluorescent calcium binding dye -- 熒光細(xì)胞成像
U2OS 細(xì)胞在 96 孔黑壁/透明底部 costar 板中以每 100 µL/孔 40,000 個細(xì)胞過夜播種。 去除生長培養(yǎng)基，將細(xì)胞與 100 µl 4 µM Fluo-4 AM 或 Fluo-8® AM 在 HHBS 中在 37 °C、5% CO2 培養(yǎng)箱中孵育 1 小時。 細(xì)胞用 200 µl HHBS 洗滌兩次，然后使用 FITC 通道在熒光顯微鏡下成像。

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