在相关光学和电子显微镜中使用抗褪色试剂进行有效的荧光回收。

IF 1.5 4区工程技术 Q3 MICROSCOPY

Microscopy Pub Date : 2019-10-09 DOI:10.1093/jmicro/dfz029

K. Toyooka, Naeko Shinozaki‐Narikawa

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引用次数: 3

摘要

相关光电子显微镜(CLEM)通过结合从荧光显微镜和电子显微镜获得的图像，使荧光标记蛋白的超微结构水平分析成为可能。CLEM方法的一个技术挑战是有效检测树脂中样品的荧光，这通常会导致荧光衰减。为了克服这个问题，我们开发了一种利用市售抗褪色试剂在树脂包埋的半薄切片中荧光恢复绿色荧光蛋白(GFP)的方法。通过应用该方法，我们成功地使用场发射扫描电镜获得了具有中等增强GFP信号的CLEM图像，证明了这种简单的荧光恢复方法的有效性。

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Efficient fluorescence recovery using antifade reagents in correlative light and electron microscopy.

Correlative light and electron microscopy (CLEM) enables ultrastructural-level analysis of fluorescence-labeled proteins by combining images obtained from both fluorescence and electron microscopies. A technical challenge with the CLEM method is the effective detection of fluorescence from samples embedded in resins, which generally cause fluorescence decay. To overcome this issue, we developed a method for fluorescence recovery of green fluorescent protein (GFP) in resin-embedded semi-thin sections using commercially available antifade reagents. By applying this method, we successfully obtained CLEM images using field-emission scanning electron microscopy with moderately enhanced GFP signals, demonstrating the efficacy of this simple fluorescence recovery method.

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来源期刊

Microscopy Physics and Astronomy-Instrumentation

CiteScore

3.30

自引率

11.10%

发文量

期刊介绍： Microscopy, previously Journal of Electron Microscopy, promotes research combined with any type of microscopy techniques, applied in life and material sciences. Microscopy is the official journal of the Japanese Society of Microscopy.