Martins Raquel R, McCracken Andrew W, Simons Mirre J P, Henriques Catarina M, Rera Michael
Department of Oncology and Metabolism, Medical School, University of Sheffield, Sheffield, UK.
Bateson Centre, University of Sheffield, Sheffield, UK.
Bio Protoc. 2018 Feb 5;8(3). doi: 10.21769/BioProtoc.2722.
The Smurf Assay (SA) was initially developed in the model organism where a dramatic increase of intestinal permeability has been shown to occur during aging (Rera , 2011). We have since validated the protocol in multiple other model organisms (Dambroise , 2016) and have utilized the assay to further our understanding of aging (Tricoire and Rera, 2015; Rera , 2018). The SA has now also been used by other labs to assess intestinal barrier permeability (Clark , 2015; Katzenberger , 2015; Barekat , 2016; Chakrabarti , 2016; Gelino , 2016). The SA in itself is simple; however, numerous small details can have a considerable impact on its experimental validity and subsequent interpretation. Here, we provide a detailed update on the SA technique and explain how to catch a Smurf while avoiding the most common experimental fallacies.
蓝精灵检测法(SA)最初是在一种模式生物中开发的,研究表明,在衰老过程中该生物的肠道通透性会显著增加(雷拉,2011年)。此后,我们在多种其他模式生物中验证了该方案(丹布罗伊斯,2016年),并利用该检测法进一步加深了我们对衰老的理解(特里夸尔和雷拉,2015年;雷拉,2018年)。现在,其他实验室也已使用SA来评估肠道屏障通透性(克拉克,2015年;卡岑贝格尔,2015年;巴雷卡特,2016年;恰克拉巴蒂,2016年;杰利诺,2016年)。SA本身很简单;然而,许多小细节可能会对其实验有效性及后续解读产生相当大的影响。在此,我们提供SA技术的详细更新内容,并解释如何在避免最常见实验谬误的同时捕捉到一个“蓝精灵”。