9 March 2016 Highly sensitive kinesin-microtubule motility assays using SLIM
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Proceedings Volume 9718, Quantitative Phase Imaging II; 97180T (2016) https://doi.org/10.1117/12.2217109
Event: SPIE BiOS, 2016, San Francisco, California, United States
Abstract
We provide an experimental demonstration of Spatial Light Interference Microscopy (SLIM) as a tool for measuring the motion of 25 nm tubulin structures without the use of florescence labels. Compared to intensity imaging methods such as phase contrast or DIC, our imaging technique relies on the ratios of images associated with optically introduced phase shifts, thus implicitly removing background illumination. To demonstrate our new found capabilities, we characterize kinesin-based motility continuously over periods of time where fluorescence would typically photobleach. We exploit this new method to compare the motility of microtubules at low ATP concentrations, with and without the tagging proteins formerly required to perform these studies. Our preliminary results show that the tags have a non-negligible effect on the microtubule motility, slowing the process down by more than 10%.
Conference Presentation
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Mikhail Kandel, Mikhail Kandel, Kai Wen Teng, Kai Wen Teng, Paul R. Selvin, Paul R. Selvin, Gabriel Popescu, Gabriel Popescu, } "Highly sensitive kinesin-microtubule motility assays using SLIM", Proc. SPIE 9718, Quantitative Phase Imaging II, 97180T (9 March 2016); doi: 10.1117/12.2217109; https://doi.org/10.1117/12.2217109
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