Journal
NATURE PROTOCOLS
Volume 11, Issue 4, Pages 617-633Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/nprot.2016.022
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Funding
- Consejo Nacional de Ciencia y Tecnologia (CONACYT) [213546]
- European Research Council (ERC)
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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.
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