Kinesin-propelled label-free microtubules imaged with interference reflection microscopy
Kinesin-propelled label-free microtubules imaged with interference reflection microscopy
复制标题
用干涉反射显微镜成像驱动蛋白驱动的无标记微管
DOI：
10.1088/1367-2630/abb47b
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发表时间：
2020
影响因子：
3.3
通讯作者：
Hess, Henry
中科院分区：
文献类型：
--
作者：
Saper, Gadiel;Hess, Henry
干涉反射显微镜（IRM）利用表面反射光与物体反射光之间的干涉来产生对比度，可用于对纳米尺寸的物体成像，例如Mahamdeh等人所证明的附着在表面上的微管。在体外滑动运动试验和各种纳米器件中，细胞骨架细丝是由表面粘附的运动蛋白在表面以上20至50纳米处推动的。在这里，我们使用IRM成像驱动的无标记微管，并表明与罗丹明标记的微管相比，这些未标记的微管更长，移动速度更快。IRM还提供了微管与表面的距离信息，并用于跟踪微管交叉事件。最后，我们在不添加氧气清除系统的情况下进行滑动实验，结果表明，与荧光显微镜成像的罗丹明标记微管相比，用IRM测量的无标记微管的光损伤要小得多。这一结果表明，IRM可以用于未来的实验，进一步探索氧自由基对滑翔实验的影响。此外，增加的速度和长度使无标签微管成为基于分子马达和细丝的工程设备的理想选择。
Interference reflection microscopy (IRM) utilizes the interference between the light reflected from the surface and the light reflected from an object to generate contrast and can be used to image nanometer size objects, such as a microtubule adhered to a surface as demonstrated by Mahamdeh et al. In in vitro gliding motility assays and in a variety of nanodevices, cytoskeletal filaments are propelled by surface-adhered motor proteins 20 to 50 nm above the surface. Here we employ IRM to image kinesin propelled label-free microtubules and show that these unlabeled microtubules are longer and move at higher velocities compared to rhodamine-labeled microtubules. IRM also provides information about the distance of an elevated microtubule from the surface and is used by us to follow microtubules crossover events. Finally, we perform the gliding assay without adding an oxygen scavenging system and show that there is significantly less photodamage for label-free microtubules measured with IRM compared to rhodamine-labeled microtubules imaged with fluorescence microscopy. This result suggests that IRM can be used for future experiments to further explore the effect of oxygen radicals on the gliding assay. Moreover, the increased velocity and length make label-free microtubules desirable for engineered devices based on molecular motors and filaments.
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影响因子：
16.6
作者：
Keya JJ;Suzuki R;Kabir AMR;Inoue D;Asanuma H;Sada K;Hess H;Kuzuya A;Kakugo A
通讯作者：
Kakugo A
使用暗场显微镜进行体外微管动力学测定。
DOI：
10.1007/978-1-0716-0219-5_4
发表时间：
2020
期刊：
Methods in molecular biology (Clifton, N.J.)
影响因子：
--
作者：
Spector,JeffreyO;Vemu,Annapurna;Roll-Mecak,Antonina
通讯作者：
Roll-Mecak,Antonina
影响因子：
64.8
作者：
HORIO, T;HOTANI, H
通讯作者：
HOTANI, H
影响因子：
17.1
作者：
Kandel, Mikhail E.;Teng, Kai Wen;Popescu, Gabriel
通讯作者：
Popescu, Gabriel
DOI：
10.1073/pnas.1306281110
发表时间：
2013-10-08
影响因子：
11.1
作者：
Aoyama, Susumu;Shimoike, Masahiko;Hiratsuka, Yuichi
通讯作者：
Hiratsuka, Yuichi