Interferometric scattering microscopy and its combination with single-molecule fluorescence imaging.

Journal article

Interferometric scattering microscopy (iSCAT) is a light scattering-based imaging modality that offers a unique combination of imaging speed and precision for tracking nanoscopic labels and enables label-free optical sensing down to the single-molecule level. In contrast to fluorescence, iSCAT does not suffer from limitations associated with dye photochemistry and photophysics, or the requirement for fluorescent labeling. Here we present a protocol for constructing an iSCAT microscope from commercially available optical components and demonstrate its compatibility with simultaneously operating single-molecule, objective-type, total internal reflection fluorescence microscopy. Given an intermediate level of experience with optics and microscopy, for instance graduate-level familiarity with laser beam steering and optical components, this protocol can be completed in a time frame of 2 weeks.

Authors: Ortega Arroyo, J; Cole, D; Kukura, P

• Publication status: Published
• Peer review status: Peer reviewed
• Publisher: Nature
• Journal: Nature protocols
• Issue: 4
• Publication date: 2016-04-01
• DOI: 10.1038/nprot.2016.022
• ISSN: 1754-2189, 1750-2799
• Pmid: 26938114
• Language: English
• Keywords: Time Factors; Optical Imaging; Microscopy, Fluorescence; Research Support, Non-U.S. Gov't; Journal Article