Integrated multi-wavelength microscope combining TIRFM and IRM modalities for imaging cellulases and other processive enzymes
| Autor: | Daguan Nong, William O. Hancock, Charles T. Anderson, Kate L. Vasquez Kuntz, Ming Tien, Zachary K. Haviland |
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| Rok vydání: | 2021 |
| Předmět: |
Point spread function
0303 health sciences Microscope Materials science biology Multi wavelength Cellulase 01 natural sciences Reflectivity Article Atomic and Molecular Physics and Optics law.invention 010309 optics 03 medical and health sciences Quantum dot law 0103 physical sciences Fluorescence microscope biology.protein Interference reflection microscopy Biological system 030304 developmental biology Biotechnology |
| Zdroj: | Biomed Opt Express |
| ISSN: | 2156-7085 |
| Popis: | We describe a multimodal microscope for visualizing processive enzymes moving on immobilized substrates. The instrument combines interference reflection microscopy (IRM) with multi-wavelength total internal reflectance fluorescence microscopy (TIRFM). The microscope can localize quantum dots with a precision of 2.8 nm at 100 frames/s, and was used to image the dynamics of the cellulase, Cel7a interacting with surface-immobilized cellulose. The instrument, which was built with off-the-shelf components and is controlled by custom software, is suitable for tracking other degradative enzymes such as collagenases, as well as motor proteins moving along immobilized tracks. |
| Databáze: | OpenAIRE |
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