Integrated multi-wavelength microscope combining TIRFM and IRM modalities for imaging cellulases and other processive enzymes

Nong, Daguan; Haviland, Zachary K.; Kuntz, Kate Vasquez; Tien, Ming; Anderson, Charles T.; Hancock, William O.

doi:10.1364/BOE.423798

Integrated multi-wavelength microscope combining TIRFM and IRM modalities for imaging cellulases and other processive enzymes

Journal Article · Tue May 11 04:00:00 EDT 2021 · Biomedical Optics Express

DOI:https://doi.org/10.1364/BOE.423798· OSTI ID:1842364

Nong, Daguan; Haviland, Zachary K.; Kuntz, Kate Vasquez; Tien, Ming; ;

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.

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Sponsoring Organization:: USDOE

Grant/Contract Number:: SC0019065

OSTI ID:: 1842364

Alternate ID(s):: OSTI ID: 1852799
OSTI ID: 2229919

Journal Information:: Biomedical Optics Express, Journal Name: Biomedical Optics Express Journal Issue: 6 Vol. 12; ISSN 2156-7085

Publisher:: Optical Society of AmericaCopyright Statement

Country of Publication:: United States

Language:: English

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An integrated multi-wavelength microscope combining TIRFM and IRM modalities for imaging cellulases and other processive enzymes Nong, Daguan; Hancock, William O. scholarsphere https://doi.org/10.26207/0mnz-xj93	dataset	January 2021

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