Reduction of lipofuscin-like autofluorescence in fluorescently labeled tissue
Reduction of lipofuscin-like autofluorescence in fluorescently labeled tissue
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DOI：
10.1177/002215549904700601
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发表时间：
1999-06-01
影响因子：
3.2
通讯作者：
Wessendorf, MW
中科院分区：
文献类型：
--
作者：
Schnell, SA;Staines, WA;Wessendorf, MW
荧光色素脂褐素随着年龄的增长在CNS细胞的细胞质中积累。由于其宽的激发和发射光谱，脂褐素样自发荧光的存在使荧光显微镜的使用复杂化（例如，荧光逆行追踪和荧光免疫细胞化学）。在这项研究中，我们检查了几种化学处理的组织切片的能力，以减少或消除脂褐素样自发荧光，而不会对其他荧光标记产生不利影响。我们发现，1-10 mM CuSO 4在50 mM乙酸铵缓冲液（pH 5）或1%苏丹黑B（SB）在70%乙醇中减少或消除猴，人或大鼠神经组织切片中的脂褐素自发荧光。这些治疗还略微降低了免疫荧光标记和荧光逆行道示踪剂的强度。然而，这些荧光团的减少远不如脂褐素样化合物的减少那么显著。我们的结论是，治疗组织与硫酸铜或SB提供了一个合理的折衷减少脂褐素样荧光和维护特定的荧光标记。
The fluorescent pigment lipofuscin accumulates with age in the cytoplasm of cells of the CNS. Because of its broad excitation and emission spectra, the presence of lipofuscin-like autofluorescence complicates the use of fluorescence microscopy (e.g., fluorescent retrograde tract tracing and fluorescence immunocytochemistry). In this study we examined several chemical treatments of tissue sections for their ability to reduce or eliminate lipofuscin-like autofluorescence without adversely affecting other fluorescent labels. We found that 1-10 mM CuSO4 in 50 mM ammonium acetate buffer (pH 5) or 1% Sudan Black B (SB) in 70% ethanol reduced or eliminated lipofuscin autofluorescence in sections of monkey, human, or rat neural tissue. These treatments also slightly reduced the intensity of immunofluorescent labeling and fluorescent retrograde tract tracers. However, the reduction of these fluorophores was far less dramatic than that for the lipofuscin-like compound. We conclude that treatment of tissue with CuSO4 or SB provides a reasonable compromise between reduction of lipofuscin-like fluorescence and maintenance of specific fluorescent labels.