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%.
| Original language | English (US) |
|---|---|
| Title of host publication | Quantitative Phase Imaging II |
| Editors | Gabriel Popescu, YongKeun Park |
| Publisher | SPIE |
| ISBN (Electronic) | 9781628419528 |
| DOIs | |
| State | Published - Jan 1 2016 |
| Event | 2nd Conference on Quantitative Phase Imaging, QPI 2016 - San Francisco, United States Duration: Feb 14 2016 → Feb 17 2016 |
Publication series
| Name | Progress in Biomedical Optics and Imaging - Proceedings of SPIE |
|---|---|
| Volume | 9718 |
| ISSN (Print) | 1605-7422 |
Other
| Other | 2nd Conference on Quantitative Phase Imaging, QPI 2016 |
|---|---|
| Country | United States |
| City | San Francisco |
| Period | 2/14/16 → 2/17/16 |
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ASJC Scopus subject areas
- Electronic, Optical and Magnetic Materials
- Biomaterials
- Atomic and Molecular Physics, and Optics
- Radiology Nuclear Medicine and imaging
Cite this
Highly sensitive kinesin-microtubule motility assays using SLIM. / Kandel, Mikhail; Teng, Kai Wen; Selvin, Paul R.; Popescu, Gabriel.
Quantitative Phase Imaging II. ed. / Gabriel Popescu; YongKeun Park. SPIE, 2016. 97180T (Progress in Biomedical Optics and Imaging - Proceedings of SPIE; Vol. 9718).Research output: Chapter in Book/Report/Conference proceeding › Conference contribution
}
TY - GEN
T1 - Highly sensitive kinesin-microtubule motility assays using SLIM
AU - Kandel, Mikhail
AU - Teng, Kai Wen
AU - Selvin, Paul R.
AU - Popescu, Gabriel
PY - 2016/1/1
Y1 - 2016/1/1
N2 - 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%.
AB - 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%.
UR - http://www.scopus.com/inward/record.url?scp=84983001809&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84983001809&partnerID=8YFLogxK
U2 - 10.1117/12.2217109
DO - 10.1117/12.2217109
M3 - Conference contribution
AN - SCOPUS:84983001809
T3 - Progress in Biomedical Optics and Imaging - Proceedings of SPIE
BT - Quantitative Phase Imaging II
A2 - Popescu, Gabriel
A2 - Park, YongKeun
PB - SPIE
ER -