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Related Concept Videos

Correlations02:20

Correlations

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Correlation means that there is a relationship between two or more variables (such as ice cream consumption and crime), but this relationship does not necessarily imply cause and effect. When two variables are correlated, it simply means that as one variable changes, so does the other. We can measure correlation by calculating a statistic known as a correlation coefficient. A correlation coefficient is a number from -1 to +1 that indicates the strength and direction of the relationship between...
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Overview of Electron Microscopy01:25

Overview of Electron Microscopy

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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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Scanning Electron Microscopy01:07

Scanning Electron Microscopy

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A scanning electron microscope (SEM) is used to study the surface features of a sample by using an electron beam that scans the sample surface in a two-dimensional manner. Typically, areas between ~1 centimeter to 5 micrometers in width can be imaged. SEM can be used to image bacteria, viruses, tissues as well as larger samples like insects. Conventional SEM gives a magnification ranging from 20X to 30,000X and spatial resolution of 50 to 100 nanometers.
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Transmission Electron Microscopy01:15

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In 1931, physicist Ernst Ruska—building on the idea that magnetic fields can direct an electron beam just as lenses can direct a beam of light in an optical microscope—developed the first prototype of the electron microscope. This development led to the development of the field of electron microscopy. In the transmission electron microscope (TEM), electrons are produced by a hot tungsten element and accelerated by a potential difference in an electron gun, which gives them up to 400...
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Immunogold Electron Microscopy01:20

Immunogold Electron Microscopy

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Immunoelectron microscopy utilizes immunogold labeling of endogenous proteins with specific antibodies to detect and localize these proteins in cells and tissues. The procedure provides insights into the distribution and quantification of protein under different stimulation conditions offering clues about their functions. Conjugating highly electron-dense gold particles with primary or secondary antibodies allow antigen detection on and within cells, with high resolution and specificity.
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Cryo-electron Microscopy01:28

Cryo-electron Microscopy

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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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Direct Comparison of Hyperspectral Stimulated Raman Scattering and Coherent Anti-Stokes Raman Scattering Microscopy for Chemical Imaging
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Cell-Nanoparticle Interactions at (Sub)-Nanometer Resolution Analyzed by Electron Microscopy and Correlative Coherent

Jukka Saarinen1, Friederike Gütter2, Mervi Lindman3

  • 1Drug Research Program, Division of Pharmaceutical Chemistry and , University of Helsinki, Viikinkaari 5 E (PO Box 56), 00014 Helsinki, Finland.

Biotechnology Journal
|October 24, 2018
PubMed
Summary
This summary is machine-generated.

Correlative Coherent Anti-Stokes Raman Scattering and Electron Microscopy (C-CARS-EM) provides label-free, high-resolution imaging of nanoparticle uptake in cells. This technique precisely visualizes nanoparticle localization within cellular compartments, advancing drug delivery research.

Keywords:
cell imagingcellular nanoparticle uptakecoherent anti-Stokes Raman scattering (CARS) microscopydrug nanocrystalsnon-linear imaging

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Correlative Light- and Electron Microscopy Using Quantum Dot Nanoparticles
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Area of Science:

  • Nanotechnology and Materials Science
  • Cell Biology and Imaging
  • Drug Delivery Systems

Background:

  • Nanoparticles are crucial for drug delivery, necessitating advanced imaging for tracking cellular interactions.
  • Label-free imaging is highly desirable to monitor nanoparticle uptake and intracellular fate without altering cellular processes.
  • Combining complementary imaging techniques enhances the detail and accuracy of studying nanoparticle-cell interactions.

Purpose of the Study:

  • To demonstrate the synergistic use of correlative Coherent Anti-Stokes Raman Scattering and Electron Microscopy (C-CARS-EM) for label-free nanoparticle imaging.
  • To investigate the cellular uptake and intracellular localization of unlabeled drug nanocrystals using the C-CARS-EM platform.
  • To assess the capability of C-CARS-EM in providing chemically-specific and high-spatial-resolution insights into nanoparticle-cell interactions.

Main Methods:

  • Employed Coherent Anti-Stokes Raman Scattering (CARS) microscopy for chemically-specific imaging with (sub)micron resolution.
  • Utilized Transmission Electron Microscopy (TEM) for ultra-high (sub)-nanometer spatial resolution imaging.
  • Integrated CARS and TEM in a correlative workflow (C-CARS-EM) to image unlabeled drug nanocrystals within macrophage cells.

Main Results:

  • Demonstrated good colocalization between CARS signals and electron-dense nanocrystals observed in TEM images.
  • TEM images revealed subcellular localization of nanocrystals within membrane-bound vesicles, characteristic of late endosomes and phagolysosomes.
  • The C-CARS-EM approach successfully visualized nanoparticle interactions at the subcellular level with high precision.

Conclusions:

  • Correlative C-CARS-EM imaging offers a powerful label-free platform for studying nanoparticle-cell interactions.
  • This technique provides both chemical specificity and ultra-high spatial resolution, essential for detailed analysis.
  • C-CARS-EM has significant potential for advancing research in nanoparticle-based drug delivery and cellular biology.