Unbiased automated quantitation of ROS signals in live retinal neurons of Drosophila using Fiji/ImageJ.
Unbiased automated quantitation of ROS signals in live retinal neurons of Drosophila using Fiji/ImageJ.
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DOI：
10.2144/btn-2021-0006
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发表时间：
2021-08
期刊：
影响因子：
2.7
通讯作者：
中科院分区：
文献类型：
--
作者：
许多成像模块用于研究细胞过程中发生的变化。除了定性（免疫组织化学）或半定量（Western印迹）方法之外，缺乏用于检测和分析疾病生物标志物的信号强度的直接定量方法。因此，需要开发在活组织成像期间定量特定信号并消除噪声的方法。活性氧（ROS）如超氧（O2·-）自由基的增加导致生物分子的氧化损伤，从而导致氧化应激。这可以通过活组织中的二氢乙锭染色来检测，其不依赖于固定，并且有助于防止对组织的应力。然而，在活组织染色中信噪比降低。我们采用果蝇眼模型的阿尔茨海默氏病作为概念的证明，以定量活性氧在活组织中采用无偏的方法。本文提出的方法对其他活组织荧光图像具有潜在的应用。我们提出了一种成像方法和定量策略，以评估氧化应激，使用荧光探针定量超氧化物（O2·-），即活性氧在活细胞成像过程中的果蝇眼模型阿尔茨海默氏病。我们试图通过采用无偏的自动定量策略来减少活细胞成像过程中的信号噪声。在这里，我们采用Fiji/ ImageJ方法进行无偏定量。
Numerous imaging modules are utilized to study changes that occur during cellular processes. Besides qualitative (immunohistochemical) or semiquantitative (Western blot) approaches, direct quantitation method(s) for detecting and analyzing signal intensities for disease(s) biomarkers are lacking. Thus, there is a need to develop method(s) to quantitate specific signals and eliminate noise during live tissue imaging. An increase in reactive oxygen species (ROS) such as superoxide (O2•-) radicals results in oxidative damage of biomolecules, which leads to oxidative stress. This can be detected by dihydroethidium staining in live tissue(s), which does not rely on fixation and helps prevent stress on tissues. However, the signal-to-noise ratio is reduced in live tissue staining. We employ the Drosophila eye model of Alzheimer's disease as a proof of concept to quantitate ROS in live tissue by adapting an unbiased method. The method presented here has a potential application for other live tissue fluorescent images. We present an imaging method and quantification strategy to assess oxidative stress using a fluorescent probe for quantitation of superoxide (O2•-), that is, reactive oxygen species during live cell imaging in the Drosophila eye model of Alzheimer's disease. We attempted to reduce noise in signal during live cell imaging by employing an unbiased automated quantitation strategy. Here, we employed the Fiji/ ImageJ method to perform an unbiased quantitation.
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影响因子：
14.9
作者：
Larkin A;Marygold SJ;Antonazzo G;Attrill H;Dos Santos G;Garapati PV;Goodman JL;Gramates LS;Millburn G;Strelets VB;Tabone CJ;Thurmond J;FlyBase Consortium
通讯作者：
FlyBase Consortium
影响因子：
3.6
作者：
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影响因子：
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影响因子：
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作者：
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影响因子：
5.2
作者：
Sanz, Alberto;Fernandez-Ayala, Daniel J. M.;Jacobs, Howard T.
通讯作者：
Jacobs, Howard T.