| In vitro Detection of Neutrophil Traps and Post-attack Cell Wall Changes in Candida Hyphae
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Author:
Date:
2017-04-05
[Abstract] In this protocol we describe how to visualize neutrophil extracellular traps (NETs) and fungal cell wall changes in the context of the coculture of mouse neutrophils with fungal hyphae of Candida albicans. These protocols are easily adjusted to test a wide array of hypotheses related to the impact of immune cells on fungi and the cell wall, making them promising tools for exploring host-pathogen interactions during fungal infection.
[摘要] 在本协议中，我们描述了如何在小鼠嗜中性粒细胞与真菌白色念珠菌的真菌菌丝共培养的背景下观察嗜中性粒细胞胞外捕获物（NETs）和真菌细胞壁变化。 这些方案很容易调整，以测试与免疫细胞对真菌和细胞壁的影响有关的各种假设，使其成为探索真菌感染期间宿主病原体相互作用的有前景的工具。
C。白色念珠菌是一种多态性机会性酵母，嗜中性粒细胞是免疫细胞，对于这种和其他真菌病原体的防御至关重要（Brown等，2012; Lionakis和Netea，2013）。 NETs是可以针对病原体部署的潜在防御机制，并且已经表明它们优先针对诸如C的微生物细胞部署。白念珠菌菌丝太大而不能吞噬（Urban et al。，2006; Bruns等人，2010; Branzk等人，，2014; Rohm等人，2014）。 NET已经显示含有许多组分，包括髓过氧化物酶，胞外DNA和瓜氨酸化的组蛋白（Amulic等人，2012; ...
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| Immunolabeling of Proteins in situ in Escherichia coli K12 Strains
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Author:
Date:
2013-08-05
[Abstract] This protocol was developed to label proteins in bacterial cells with antibodies conjugated to a fluorophore for fluorescence microscopy imaging. The procedure is optimized to minimize morphological changes and also to minimize the amount of antibodies needed for the staining. The protocol can also be used with primary antibodies conjugated to a fluorophore. The method has been verified extensively (van der Ploeg et al., 2013), but it should be noted that one case in Caulobacter crescentus (Hocking et al., 2012) has been reported in which the localization of a protein changed upon fixation by formaldehyde/glutaraldehyde. However, the localization of the same protein in E. coli did not change.
[摘要] 该协议被开发来标记细菌细胞中的蛋白质与缀合到荧光显微镜成像荧光的抗体。 优化程序以使形态变化最小化，并且使染色所需的抗体的量最小化。 该方案还可以与结合至荧光团的一抗一起使用。 该方法已经广泛地验证（van der Ploeg等人，2013），但是应当注意的是，在新月柄杆菌中的一种情况（Hocking等人，/2012>），其中蛋白质的定位在通过甲醛/戊二醛固定时改变。 然而，相同蛋白质在E中的定位。 大肠杆菌没有改变。
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