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Related Concept Videos

Overview of Microscopy Techniques01:22

Overview of Microscopy Techniques

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The early pioneers of microscopy opened a window into the invisible world of microorganisms. In 1830, Joseph Jackson Lister created an essentially modern light microscope. The 20th century saw the development of microscopes that leveraged nonvisible light, such as fluorescence microscopy that uses an ultraviolet light source and electron microscopy that uses short-wavelength electron beams. These advances significantly improved magnification, image resolution, and contrast. By comparison, the...
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Electron Microscope Tomography and Single-particle Reconstruction01:07

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Transmission electron microscopy (TEM) can be used to determine the 3D structure of biological samples with the help of techniques such as electron microscope tomography and single-particle reconstruction. While single-particle reconstruction can examine macromolecules and macromolecular complexes in vitro conditions only, tomography permits the study of cell components or small cells in vivo.
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Three-Dimensional Microscopy in Microbiology01:28

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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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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...
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Conventional electron microscopy (EM) involves dehydration, fixation, and staining of biological samples, which distorts the native state of biological molecules and results in several artifacts. Also, the high-energy electron beam damages the sample and makes it difficult to obtain high-resolution images. These issues can be addressed using cryo-EM, which uses frozen samples and gentler electron beams. The technique was developed by Jacques Dubochet, Joachim Frank, and Richard Henderson, for...
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Overview of Electron Microscopy01:25

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The wavelengths of visible light ultimately limit the maximum theoretical resolution of images created by light microscopes. Most light microscopes can only magnify 1000X, and a few can magnify up to 1500X. Electrons, like electromagnetic radiation, can behave like waves, but with wavelengths of 0.005 nm, they produce significantly greater resolution up to 0.05 nm as compared to 500 nm for visible light. An electron microscope (EM) can create a sharp image that is magnified up to 2,000,000X.
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Microscopic Imaging Techniques for Molecular Assemblies: Electron, Atomic Force, and Confocal Microscopies.

Ryou Kubota1, Wataru Tanaka1, Itaru Hamachi1,2

  • 1Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Katsura, Nishikyo-ku, Kyoto 615-8510, Japan.

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|May 4, 2021
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Summary

Microscopic imaging visualizes molecular assemblies, aiding the understanding of structure-function relationships for designing advanced materials. This review covers various imaging techniques and applications for self-assembled structures.

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Area of Science:

  • Supramolecular chemistry and materials science.
  • Advanced microscopic imaging techniques.

Background:

  • Self-assembly is crucial for creating diverse supramolecular structures with functions dependent on their architecture.
  • Understanding structure-function relationships is key for designing functional molecular assemblies.
  • Microscopy offers high-resolution structural insights complementary to ensemble spectroscopic methods.

Purpose of the Study:

  • To review representative examples of molecular assembly visualization using various microscopy techniques.
  • To discuss advanced imaging methods for dynamic and complex self-assembly processes.
  • To provide guidelines for structural analysis to advance the development of next-generation materials.

Main Methods:

  • Electron microscopy (EM).
  • Atomic force microscopy (AFM).
  • Confocal microscopy.
  • Super-resolution microscopy.

Main Results:

  • Detailed visualization of supramolecular nanofibers, gels, micelles, vesicles, coacervate droplets, polymer assemblies, and protein/DNA assemblies.
  • Demonstration of advanced imaging capabilities for evaluating assembly dynamics, multicomponent systems, and in-cell processes.

Conclusions:

  • Microscopic imaging is a powerful tool for elucidating the structure of molecular assemblies.
  • Advanced imaging techniques enable deeper structural analyses and understanding of complex self-assembly.
  • This review guides the rational design and development of novel functional molecular materials.