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在此方案中,全细胞膜片钳记录由神经元细胞使用含 50 µM Alexa Fluor® 633 的细胞内记录液获得。因此,仅记录的神经元采用荧光染料灌注,随后用该方案处理切片后在共聚焦显微镜下进行观察。
In this protocol, whole cell patch-clamp recordings were made from neurons using an intracellular recording solution containing 50 µM Alexa Fluor® 633. Thus, only the recorded neurone will be filled with a fluorescent dye that will subsequently be visualized on the confocal microscope following the treatment of the slice with this protocol.
将脑片从记录室中取出,在 pH 为 7.4 且含 4% PFA(多聚甲醛)的 0.1 M 磷酸盐缓冲液中浸没过夜。
第 1 天
第 2 天
重复此步骤 2 次,直到您已冲洗 3 次。
放置盖玻片后,立即使用共聚焦显微镜观察样本。
视频方案由 Abcam 与NeuroSolutions合作制作。