What is structured illumination?
Structured Illumination Microscopy (SIM) is a super-resolution fluorescence optical microscope imaging technique that increases resolution by exploiting interference patterns (moiré patterns) created when two grids are overlaid at an angle.
What is structured illumination microscopy used for?
Structured illumination is used to enhance spatial resolution and involves illuminating the sample with patterned light and using software to analyze the information in Moiré fringes outside the normal range of observation.
How does 3D SIM work?
3D SIM is a wide-field imaging technique in which the entire field of view is illuminated with a striped pattern of interfering light know as Moiré fringes. This grid shadow is typically projected onto the specimen and only become visible in the focal plane.
How does expansion microscopy work?
Expansion microscopy is achieved by synthesizing a polymer system within a specimen. By then swelling this polymer network, the sample is expanded to be examined under conventional microscopic analysis tools without degrading the integrity of the sample.
What’s the meaning of deconvolution?
Definition of deconvolution : simplification of a complex signal (as instrumental data) usually by removal of instrument noise.
What is the advantage of Köhler illumination?
The primary advantage of Köhler illumination is the uniform illumination of the sample. This reduces image artifacts and provides high sample contrast. Uniform illumination of the sample is also critical for advanced illumination techniques such as phase contrast and differential interference contrast microscopy.
Does Köhler illumination increase magnification?
Resolution and contrast, however, are largely dependent on specimen illumination. Optical illumination for most specimens is called Kohler Illumination. Once a given selection of optics is in place, Kohler illumination cannot affect magnification, but it does affect resolution and contrast.
What is structured illumination and how does it work?
Structured illumination utilizes specific patterns of light to determine the geometric shape and depth of objects. An effective 3D system can be constructed by illuminating objects with different patterns, such as grids, dots, or lines, while minimizing cost, components, and complexity.
What is structured illumination microscopy?
Structured illumination microscopy is a technique for acquiring lateral- and axial-resolution optical images under the abovementioned diffraction limit using sets of sinusoidal-patterned illumination and post-processing to extract high-frequency components [40–43].
What is structured illumination in LSM?
Structured illumination in LSM is typically used for contrast enhancement rather than an improvement in resolution. This is because the lateral resolution improvement achieved by SIM is dependent on the spatial frequency of the illumination grid and the maximum spatial frequency, which can be resolved by the detection optics.
Why is it important to calibrate structured illumination?
Since a well-calibrated system increases measurement accuracy, it is important to understand that structured illumination is not universal, and certain structures should be used to obtain certain measurements.