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Related Concept Videos

Imaging Biological Samples with Optical Microscopy01:18

Imaging Biological Samples with Optical Microscopy

Optical microscopy uses optic principles to provide detailed images of samples. Antonie van Leeuwenhoek designed the first compound optical microscope in the 17th century to visualize blood cells, bacteria, and yeast cells. In 1830, Joseph Jackson Lister created an essentially modern light microscope. The 20th century saw the development of microscopes with enhanced magnification and resolution.
In optical microscopy, the specimen to be viewed is placed on a glass slide and clipped on the stage...
Super-resolution Fluorescence Microscopy01:37

Super-resolution Fluorescence Microscopy

Super-resolution fluorescence microscopy (SRFM) provides a better resolution than conventional fluorescence microscopy by reducing the point spread function (PSF). PSF is the light intensity distribution from a point that causes it to appear blurred. Due to PSF, each fluorescing point appears bigger than its actual size, and it is the PSF interference of nearby fluorophores that causes the blurred image. Various approaches to achieving higher resolution through SRFM have recently been developed.
Confocal Fluorescence Microscopy01:16

Confocal Fluorescence Microscopy

Confocal microscopy is an advanced microscopic technique. The prime advantage of the confocal microscope over other microscopy techniques is its ability to block the out-of-focus light from the illuminated samples using pinholes. It is widely used with fluorescence optics to obtain high-resolution, sharp contrast images. Unlike optical microscopes, confocal microscopes use a focused beam of light laser to scan the entire sample surface at different z-planes. These microscopes are, therefore,...
Phase Contrast and Differential Interference Contrast Microscopy01:26

Phase Contrast and Differential Interference Contrast Microscopy

Phase-Contrast Microscopes
In-phase-contrast microscopes, interference between light directly passing through a cell and light refracted by cellular components is used to create high-contrast, high-resolution images without staining. It is the oldest and simplest type of microscope that creates an image by altering the wavelengths of light rays passing through the specimen. Altered wavelength paths are created using an annular stop in the condenser. The annular stop produces a hollow cone of...
Three-Dimensional Microscopy in Microbiology01:28

Three-Dimensional Microscopy in Microbiology

Three-dimensional imaging techniques are essential in cell biology, allowing researchers to visualize intricate cellular structures with high resolution. Two prominent methods, Differential Interference Contrast Microscopy (DIC) and Confocal Scanning Laser Microscopy (CSLM), provide distinct advantages for imaging live and thick specimens, respectively.Differential Interference Contrast MicroscopyDIC microscopy enhances contrast in transparent, unstained samples by converting phase...

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A Guide to Structured Illumination TIRF Microscopy at High Speed with Multiple Colors
11:15

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Published on: May 30, 2016

Optimization and characterization of a structured illumination microscope.

Frédéric Chasles, Benoit Dubertret, A Claude Boccara

    Optics Express
    |June 25, 2009
    PubMed
    Summary
    This summary is machine-generated.

    Structured illumination microscopy offers a cost-effective way to achieve optical sectioning, improving axial resolution by 1.5x. This scanning-free technique enhances fluorescence microscopy performance for detailed sample imaging.

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    Cryo-Structured Illumination Microscopic Data Collection from Cryogenically Preserved Cells

    Published on: May 28, 2021

    Area of Science:

    • Biophotonics
    • Microscopy techniques
    • Optical imaging

    Background:

    • Structured illumination microscopy (SIM) is a cost-effective method for optical sectioning.
    • It serves as a scanning-free alternative to confocal microscopy for fluorescent samples.
    • Regular fluorescent microscopes can be easily adapted for SIM implementation.

    Purpose of the Study:

    • To theoretically analyze SIM performance, including sectioning strength, axial resolution, and signal-to-background ratio.
    • To evaluate the impact of objective lens and grid characteristics (pitch, contrast) on SIM performance.
    • To compare SIM with commercial microscopy setups like confocal, Spinning Disk, and Zeiss Apotome.

    Main Methods:

    • Theoretical analysis of SIM performance metrics.
    • Modification of the original grid in-step modulation to sinusoidal modulation.
    • Computation of optical sections using four images acquired during one modulation period.
    • Comparison of the developed SIM technique with commercial confocal, Spinning Disk, and Zeiss Apotome systems.

    Main Results:

    • Optimized SIM conditions can improve axial resolution by 1.5x compared to epifluorescence microscopy.
    • Optical sections with a thickness below 400nm are achievable with a 1.4 numerical aperture objective.
    • The developed sinusoidal modulation algorithm is stable at high frequencies, with acquisition speed limited by detector performance and signal-to-background ratio.

    Conclusions:

    • Structured illumination microscopy provides a simple, cheap, and effective method for optical sectioning with enhanced axial resolution.
    • The technique offers a viable scanning-free alternative to confocal microscopy for fluorescence imaging.
    • The developed algorithm demonstrates robustness and efficiency, comparable to commercial advanced microscopy systems.