Kural Comert, Nonet Michael L, Selvin Paul R
Center for Biophysics and Computational Biology, University of Illinois, Urbana, Illinois 61801, USA.
Biochemistry. 2009 Jun 9;48(22):4663-5. doi: 10.1021/bi8023477.
Despite much work, subcellular neurons of Caenorhabditis elegans have not been studied at nanometer resolution with millisecond time resolution. Nor has there been an effective way to immobilize C. elegans. Here we show that, without using anesthetic or paralyzing agents, fluorescence imaging with one-nanometer accuracy (FIONA) can be successfully applied to fluorescently labeled molecules within C. elegans nerves. GFP- and DENDRA2-labeled ELKS punctae can be localized with sub-10 nm accuracy in approximately 5 ms. Our results show that the protein ELKS is occasionally transferred by microtubule-based motors. This is the first example of FIONA applied to a living organism.
尽管已经开展了大量工作,但秀丽隐杆线虫的亚细胞神经元尚未在具有毫秒级时间分辨率的纳米分辨率下进行研究。而且也没有一种有效的方法来固定秀丽隐杆线虫。在此我们表明,在不使用麻醉剂或麻痹剂的情况下,具有一纳米精度的荧光成像(FIONA)能够成功应用于秀丽隐杆线虫神经内的荧光标记分子。绿色荧光蛋白(GFP)和德伦拉2(DENDRA2)标记的ELKS点状结构可在大约5毫秒内以低于10纳米的精度进行定位。我们的结果表明,ELKS蛋白偶尔会由基于微管的马达转运。这是FIONA应用于活体生物的首个实例。