| Non-separate Mouse Sclerochoroid/RPE/Retina Staining and Whole Mount for the Integral Observation of Subretinal Layer
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Author:
Date:
2021-01-05
[Abstract] The subretinal layer between retinal pigment epithelium (RPE) and photoreceptors is a region involved in inflammation and angiogenesis during the procession of diseases such as age-related macular degeneration. The current protocols of whole mounts (retina and RPE) are unable to address the intact view of the subretinal layer because the separation between retina and RPE is required, and each separate tissue is then stained. Non-separate Sclerochoroid/RPE/Retina whole mount staining was recently developed and reported. The method can be further combined and optimized with melanin bleaching and tissue clearing. Here, we describe steps of both non-pigmented and pigmented mouse Sclerochoroid/RPE/Retina whole mount including eyeball preparation, staining, mounting and confocal scanning. In ...
[摘要] [摘要]视网膜色素上皮（RPE）和感光细胞之间的视网膜下层是与年龄相关的黄斑变性等疾病进程中涉及炎症和血管生成的区域。由于需要在视网膜和RPE之间分离，因此整个安装（视网膜和RPE）的当前方案无法解决视网膜下层的完整视图，然后对每个单独的组织进行染色。最近开发并报道了非分离的巩膜脉络膜/ RPE /视网膜整装染色。该方法可以与黑色素漂白和组织清除进一步结合和优化。在这里，我们描述了非色素和色素小鼠硬化脉络膜的步骤/ RPE /视网膜整个安装，包括眼球准备，染色，安装和共聚焦扫描。此外，我们提出了巩膜脉络膜/ RPE /视网膜整个支架中的RPE，视网膜下小胶质细胞和邻近的感光体的共聚焦图像。
[背景]在眼睛视网膜是由视网膜色素上皮（RPE）包围，脉络膜和巩膜。通常，在整装染色中，视网膜组织与脉络膜/ RPE分开，并且分开的视网膜和脉络膜/ RPE的每个部分都被染色。因此，视网膜或脉络膜/ RPE的单独的整体染色不能解决完整的视网膜下信息。最近，开发了脉络膜/ RP E /视网膜整个安装方案（Kim等，2016； ...
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| Measurement of Intracellular ROS in Caenorhabditis elegans Using 2’,7’-Dichlorodihydrofluorescein Diacetate
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Author:
Date:
2018-03-20
[Abstract] Reactive oxygen species (ROS) are generated during normal metabolic processes under aerobic conditions. Since ROS production initiates harmful radical chain reactions on cellular macromolecules, including lipid peroxidation, DNA mutation, and protein denaturation, it has been implicated in a wide spectrum of diseases such as cancer, cardiovascular disease, ischemia-reperfusion and aging. Over the past several decades, antioxidants have received explosive attention regarding their protective potential against these deleterious reactions. Accordingly, many analytical methodologies have been developed for the evaluation of the antioxidant capacity of compounds or complex biological samples. Herein, we introduce a simple and convenient method to detect in vivo intracellular ROS ...
[摘要] 在有氧条件下的正常代谢过程中产生活性氧物质（ROS）。 由于ROS产生会引起细胞大分子的有害自由基连锁反应，包括脂质过氧化反应，DNA突变和蛋白质变性，因此它已涉及多种疾病，如癌症，心血管疾病，缺血再灌注和衰老。 在过去的几十年中，抗氧化剂已经受到爆炸性的关注，因为它们对这些有害反应具有保护作用。 因此，已经开发了许多分析方法用于评估化合物或复杂生物样品的抗氧化能力。 在此，我们介绍了使用2'，7'-二氯荧光素二乙酸盐（H 2），在线虫体内用光度计检测体内细胞内ROS水平的简单方便的方法。 DCFDA），一种细胞渗透性示踪剂。
【背景】已经广泛地使用荧光探针2'，7'-二氯荧光素二乙酸酯（H 2 DCFDA）原位检测活体内细胞内活性氧簇（ROS）水平由在氧化应激和相关疾病领域工作的研究人员提供。非极性和非离子型探针H 2 DCFDA可以容易地穿透细胞膜并且被酯酶酶促脱乙酰化。该生物化学反应将H 2 DCFDA转化成非荧光化合物H 2 DCF，然后将其迅速氧化成高度荧光的2'，7'-二氯荧光素（DCF） ROS的存在（图1A）。因此，来自H 2 ...
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| Nictation Assays for Caenorhabditis and Other Nematodes
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Author:
Date:
2015-04-05
[Abstract] Nictation is a dauer-specific standing and waving behavior of the nematodes including Caenorhabditis species. Nictation enhances the capability of free-living nematodes to hitchhike to other animals as well as parasitic nematodes to infect their hosts. However, lack of an assay for this behavior has made it difficult to elucidate its underlying regulatory mechanisms and related genetic pathways. We have developed nictation assays that enable the quantification of the nictation behavior of individuals and groups of worms. Gauze assay is less quantitative but is an easier way to observe nictation behavior in plates with plenty of dauers. The micro-dirt chip made from PDMS mold is a more sophisticated method to quantify the nictation behavior. Nictation can be quantified on a ...
[摘要] 捕食是线虫（包括Caenorhabditis）物种的持久性和持续性行为。 捕食提高了自由生活线虫与其他动物以及寄生线虫感染其宿主的能力。 然而，缺乏这种行为的测定使得难以阐明其基础调节机制和相关的遗传途径。 我们已经开发了定义测定，使得能够量化的个人和蠕虫组的nictation行为。 纱布测定较少定量，但是是更容易的方法来观察具有大量持久的板的行为。 由PDMS模具制成的微尘片是一种更复杂的方法来量化叙述行为。 可以通过测量个体动物的平均哺乳时间或通过在给定动物群体中的ating虫的分数，在微尘片上量化捕食。
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