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Related Concept Videos

Studying the Cytoskeleton01:17

Studying the Cytoskeleton

The cytoskeletal architecture can be studied using different microscopic and biochemical techniques. Electron microscopy was instrumental in discovering the cytoskeletal architecture around the 1960s, which allowed obtaining structural information at a high-resolution level. However, the sample preparation procedure often limits this ability in biological samples. Several protocols have been developed over the years to optimize sample preparation. In one of the protocols known as rotary...
Overview of Electron Microscopy01:25

Overview of Electron Microscopy

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.
Preparation of Samples for Electron Microscopy01:20

Preparation of Samples for Electron Microscopy

To be visualized by an electron microscope, either transmission or scanning, biological samples need to be fixed (stabilized) so the electron beam does not destroy them and dried thoroughly (desiccated/dehydrated) so the vacuum does not affect them. Fixation needs to be done as quickly as possible because the sample properties will start changing as soon as it is removed from its natural environment. For example, in a tissue sample, the oxygen levels begin decreasing, causing an altered...
Immunogold Electron Microscopy01:20

Immunogold Electron Microscopy

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.
Cryo-electron Microscopy01:28

Cryo-electron Microscopy

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...
Electron Microscope Tomography and Single-particle Reconstruction01:07

Electron Microscope Tomography and Single-particle Reconstruction

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.
Electron Tomography
Electron tomography can be performed either in TEM or STEM (scanning transmission...

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Correlative Light- and Electron Microscopy Using Quantum Dot Nanoparticles
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Electron microscopic examination of cytologic samples

E Ito1, H Nei, M Noda

  • 1Department of Obstetrics and Gynecology, School of Medicine, Sapporo Medical University, Japan.

Acta Cytologica
|October 2, 1998
PubMed
Summary

Successive examination using light microscopy (LM), scanning electron microscopy (SEM), and transmission electron microscopy (TEM) enhances cell identification. Electron microscopy (EM) findings are valuable for cytodiagnosis when LM is insufficient.

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Area of Science:

  • Cytopathology
  • Electron Microscopy
  • Cell Biology

Background:

  • Light microscopy (LM) is standard for cytodiagnosis.
  • Limitations exist in LM for definitive cell identification and origin determination.
  • Advanced microscopy techniques may offer complementary diagnostic information.

Purpose of the Study:

  • To evaluate the diagnostic utility of sequential light, scanning electron, and transmission electron microscopy (LM-SEM-TEM) on the same cytologic sample.
  • To assess the ability of SEM and TEM to determine cell origin and malignancy.
  • To determine the added value of electron microscopy in cytodiagnosis.

Main Methods:

  • A sequential LM-SEM-TEM examination method was applied.
  • 201 cytologic specimens were analyzed over a seven-year period (1986-1993).
  • The study investigated estimation of histologic origin and malignancy from SEM and TEM findings.

Main Results:

  • Sequential LM-SEM-TEM allowed for basic interpretations of cellular ultrastructure.
  • Electron microscopy findings aided in determining cell biologic characteristics and tissue origin when LM was inconclusive.
  • Electron microscopy provided crucial information for improving cytodiagnosis.

Conclusions:

  • SEM and/or TEM findings are valuable for characterizing cells when LM is insufficient.
  • Electron microscopy contributes to understanding cellular morphology and biology.
  • Continued data accumulation in EM of cytologic samples is expected to enhance LM-based cytodiagnosis accuracy.