Improved fluorescent signal in expansion microscopy using fluorescent Fab fragment secondary antibodies.
Improved fluorescent signal in expansion microscopy using fluorescent Fab fragment secondary antibodies.
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DOI：
10.1016/j.mex.2022.101796
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发表时间：
2022
期刊：
影响因子：
1.9
通讯作者：
Stiles, Megan A.
中科院分区：
文献类型：
--
作者：
Sherry, David M.;Stiles, Megan A.
扩展显微镜（ExM）是一种显微成像方法，可以实现超分辨率的可视化荧光标记的生物样品使用常规荧光显微镜。该方法是基于将荧光标记的生物样品嵌入水凝胶基质中，然后对样品进行物理膨胀，然后使用常规荧光显微镜进行观察。该方法的变体可用于可视化内源性表达的荧光蛋白，如GFP、荧光标记的抗体、核酸或其他荧光标记的分子。该方法的一个重大挑战是，试样的物理膨胀产生伴随的荧光强度降低，这可能使成像困难。我们描述了一种放大荧光信号的方法，该方法通过应用荧光标记的Fab片段二抗来增强荧光信号，并增强使用常规荧光显微镜对标记的检测。本文描述了一种提高膨胀显微镜标本免疫荧光信号强度的方法。方法采用市售试剂。增强在扩展组织样本中获取有用图像的能力。膨胀显微镜（ExM）方法可以导致荧光强度的损失，试样的膨胀。扩增后的ExM程序中使用Fab片段二抗可以提高荧光强度。
Expansion microscopy (ExM) is a microscopic imaging approach that can achieve super-resolution visualization of fluorescently labeled biological samples using conventional fluorescence microscopy. The method is based on embedding of a fluorescently labeled biological sample in a hydrogel matrix followed by the physical expansion of the specimen, which is then viewed using a conventional fluorescent microscope. Variations of the method can be used to visualize endogenously expressed fluorescent proteins, such as GFP, fluorescently tagged antibodies, nucleic acids, or other fluorescently tagged molecules. A significant challenge of the method is that the physical expansion of the specimen produces a concommitant reduction in fluorescence intensity, which can make imaging difficult. We describe an approach for amplifying fluorescence signal following expansion of immunolabeled tissue sections by applying fluorescently labeled Fab fragment secondary antibodies to intensify fluorescent signal and enhance detection of labeling using conventional fluorescent microscopy. A method to increase immunofluorescence signal intensity of Expansion Microscopy specimens is described. Method utilizes commercially available reagents. Enhances ability to acquire useful images in expanded tissue samples. Expansion Microscopy (ExM) methods can result in loss of fluorescence intensity upon expansion of the specimen. Use of Fab fragment secondary antibodies in post-expansion ExM procedures can improve fluorescence intensity.
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光学成像。膨胀显微镜。
DOI：
10.1126/science.1260088
发表时间：
2015-01-30
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Chen F;Tillberg PW;Boyden ES
通讯作者：
Boyden ES
影响因子：
7.7
作者：
Truckenbrodt S;Maidorn M;Crzan D;Wildhagen H;Kabatas S;Rizzoli SO
通讯作者：
Rizzoli SO
影响因子：
--
作者：
Asano, Shoh M;Gao, Ruixuan;Boyden, Edward S
通讯作者：
Boyden, Edward S
Click-ExM 可对所有生物分子进行扩展显微镜检查
影响因子：
48
作者：
Sun, De-en;Fan, Xinqi;Chen, Xing
通讯作者：
Chen, Xing
影响因子：
3.8
作者：
Azuma, Takuya;Kei, Takayuki
通讯作者：
Kei, Takayuki