An integrated multi-wavelength microscope combining TIRFM and IRM modalities for imaging cellulases and other processive enzymes
Abstract: 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.
|Work Title||An integrated multi-wavelength microscope combining TIRFM and IRM modalities for imaging cellulases and other processive enzymes|
|License||CC BY-NC-ND 4.0 (Attribution-NonCommercial-NoDerivatives)|
|Deposited||April 30, 2021|
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