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

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 (TIRFM). can localize quantum dots precision of 2.8 nm at 100 frames/s, and was used to image the dynamics cellulase, Cel7a interacting surface-immobilized cellulose. instrument, which built off-the-shelf components is controlled by custom software, suitable tracking other degradative such as collagenases, well motor proteins along tracks.