Interferometric Scattering Microscopy
Interferometric Scattering Microscopy - Related Videos
11:57
Three-dimensional Super Resolution Microscopy of F-actin Filaments by Interferometric PhotoActivated Localization Microscopy (iPALM)
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Cited by 10 •
2016
We present a protocol for the application of interferometric PhotoActivated Localization Microscopy (iPALM), a 3-dimensional single-molecule localization super resolution microscopy method, to the imaging of the actin cytoskeleton in adherent mammalian cells. This approach allows light-based visualization of nanoscale structural features that would otherwise remain unresolved by conventional diffraction-limited optical microscopy.
14:58
Optical Scatter Microscopy Based on Two-Dimensional Gabor Filters
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2010
We demonstrate a dark-field microscopy method based on Gabor-like filtering to measure subcellular dynamics within single living cells. The technique is sensitive to alterations in the structure of organelles, such as mitochondrial fragmentation.
11:04
Biomolecular Detection employing the Interferometric Reflectance Imaging Sensor (IRIS)
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Cited by 2 •
2011
Quantitative, high-throughput, real-time, and label-free biomolecular detection (DNA, protein, etc.) on SiO2 surfaces can be achieved using a simple interferometric technique which relies on LED illumination, minimal optical components, and a camera. The Interferometric Reflectance Imaging Sensor (IRIS) is inexpensive, simple to use, and amenable to microarray formats.
10:16
Digital Inline Holographic Microscopy (DIHM) of Weakly-scattering Subjects
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Cited by 9 •
2014
The three-dimensional locations of weakly-scattering objects can be uniquely identified using digital inline holographic microscopy (DIHM), which involves a minor modification to a standard microscope. Our software uses a simple imaging heuristic coupled with Rayleigh-Sommerfeld back-propagation to yield the three-dimensional position and geometry of a microscopic phase object.
15:06
Measurement of Scattering Nonlinearities from a Single Plasmonic Nanoparticle
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Cited by 2 •
2016
Saturable and reverse saturable scattering were discovered in isolated plasmonic particles and adopted as a novel non-bleaching contrast method in super-resolution microscopy. Here the experimental procedures of detecting and extracting nonlinear scattering are explained in detail, as well as how to enhance resolution with the aid of saturated excitation microscopy.
FAQs
The measured signal arises from the combined optical fields, not from scattered light alone. Light scattered by a nanoscale object interferes with light reflected from the coverslip, producing an intensity contrast. Because this contrast is linked to the object’s polarizability, size, and refractive index, the measurement can reveal changes in a particle’s optical response during biochemical events.
Polarizability, size, and refractive index are the principal factors connecting an object to its observed contrast. A change in any of these properties can alter the measured intensity, so contrast should be interpreted as an optical signature rather than as a direct, isolated measurement of size alone. This relationship helps when comparing biomolecular particles or tracking their changes over time.
Unlike approaches that require fluorescent tags, Interferometric Scattering Microscopy observes the scattered-light response of the object itself. Avoiding a label can reduce perturbation to the biomolecular system while retaining sensitivity to nanoscale structures. That combination is particularly useful when tagging could influence binding, assembly, transport, or other behavior being examined.
A basic measurement begins with illumination of the sample in a configuration that provides a coverslip reflection as the reference field. The instrument then detects the intensity pattern produced by interference between that reflection and light scattered by an object. Monitoring this contrast over time supports real-time observation of individual nanoscale biological structures.
In biochemical experiments, changes in the interference contrast can be followed as molecules bind, assemble, or move through a system. The same label-free readout can support mass-related measurements, because the optical signal depends on particle properties that include size and refractive index. These capabilities let investigators connect nanoscale observations with evolving biomolecular behavior.
The method can be applied to single proteins, viruses, lipid vesicles, and other small biological structures. This range spans molecular species and larger assemblies, allowing biochemists to examine how nanoscale entities appear and change without adding fluorescent tags. Its value is therefore not limited to one class of sample, but extends across distinct biomolecular systems and their interactions.