Related Experiment Video
Updated: Aug 12, 2026
08:04
Preparation and Observation of Thick Biological Samples by Scanning Transmission Electron Tomography
Published on: March 12, 2017
A method for the examination of the same cell using light, scanning and transmission electron microscopy
1Electron Microscope Unit, Prince Henry Hospital, Sydney NSW, Australia.
Summary
This study introduces a novel method to prepare single cells for detailed analysis using light, scanning electron, and transmission electron microscopy. This technique enables comparative investigation of the same cell across multiple microscopy techniques for enhanced cytological diagnosis.
Area of Science:
- Cell Biology
- Microscopy Techniques
- Cytology
Background:
- Comparative analysis of cells using different microscopy techniques is challenging.
- Locating the exact same cell across multiple imaging modes requires precise methods.
Purpose of the Study:
- To develop a method for preparing a single cell for comparative analysis using light microscopy, scanning electron microscopy, and transmission electron microscopy.
- To facilitate the precise relocation of cells for multi-modal investigation.
Main Methods:
- A cytospin preparation was used as the starting material.
- A permanent, numbered grid pattern was etched onto glass microscope slides.
- The grid pattern allowed for consistent cell location across different microscopy techniques.
Main Results:
- The described method enables the investigation of the same cell or cell group using three distinct microscopy modes.
- Data acquisition from a single cell was achieved from light microscopy, scanning electron microscopy, and transmission electron microscopy.
Conclusions:
- This method serves as a valuable addition to routine cytological diagnostic procedures.
- The technique enhances the depth of cellular investigation by enabling multi-modal microscopic analysis of individual cells.
Related Concept Videos
01:25Overview 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.
01:07Scanning Electron Microscopy
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.
Fundamental Principles
Accelerated...
Fundamental Principles
Accelerated...
01:15Transmission Electron Microscopy
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 keV in...
01:20Immunogold 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.
01:07Electron 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...
Electron Tomography
Electron tomography can be performed either in TEM or STEM (scanning transmission...
01:22Overview of Microscopy Techniques
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...