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.

Citation

Nong, Daguan; HANCOCK, WILLIAM O (2021). An integrated multi-wavelength microscope combining TIRFM and IRM modalities for imaging cellulases and other processive enzymes [Data set]. Scholarsphere. https://doi.org/10.26207/0mnz-xj93

Files

Metadata

Work Title An integrated multi-wavelength microscope combining TIRFM and IRM modalities for imaging cellulases and other processive enzymes
Access
Open Access
Creators
  1. Daguan Nong
  2. WILLIAM O HANCOCK
Keyword
  1. single-molecule imaging,TIRFM,IRM,multi-wavelength,micromirror
License CC BY-NC-ND 4.0 (Attribution-NonCommercial-NoDerivatives)
Work Type Dataset
Acknowledgments
  1. The authors thank Dannielle Gibson and Scott Pflumm for efforts in initiating the cellulase project, and Thomas Starr for technical advice. The authors also thank the Penn State CSL Behring Fermentation Facility - University Park, PA for assistance with cellulose preparation
Publication Date 2021
DOI doi:10.26207/0mnz-xj93
Deposited April 30, 2021

Versions

Analytics

Collections

This resource is currently not in any collection.

Work History

Version 1
published

  • Created
  • Updated Acknowledgments Show Changes
    Acknowledgments
    • The authors thank Dannielle Gibson and Scott Pflumm for efforts in initiating the cellulase project, and Thomas Starr for technical advice. The authors also thank the Penn State CSL Behring Fermentation Facility - University Park, PA for assistance with cellulose preparation
  • Added Creator Daguan Nong
  • Added Creator WILLIAM O HANCOCK
  • Added Fig3a_QPD_ZPOS_Calibration.m
  • Added Fig3c_StepResponse.m
  • Added Fig4b_PixelToPhysicCalibration.m
  • Added Fig4c_QdotStepwise.m
  • Added Fig5b_s4bAndIRM.m
  • Added Fig6b_PlotScatterXY.m
  • Added Fig6c_PlotDistance.m
  • Added pixel_PosY_calibration.mat
  • Added Pixel_XPos_Calbration.mat
  • Added ProcesiveMove.mat
  • Added ProcesiveMove1.mat
  • Added QdotStepwiseMovment.mat
  • Added RawDataForQpdY-Zpos_Calibration.mat
  • Added s4bAndIRMIntensity.mat
  • Added TIRFLockedStepResponceStagesInfo.mat
  • Added Fig5a_left.tif
  • Added Fig5a_right.tif
  • Added Fig5c.tif
  • Updated License Show Changes
    License
    • https://www.gnu.org/licenses/gpl.html
  • Published
  • Updated
  • Updated

Version 2
published

  • Created
  • Updated License, Publication Date Show Changes
    License
    • https://www.gnu.org/licenses/gpl.html
    • https://creativecommons.org/licenses/by-nc-nd/4.0/
    Publication Date
    • 2012
    • 2021
  • Published
  • Updated
  • Updated

Version 3
published

  • Created
  • Added Readme.md
  • Published
  • Updated
  • Updated