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Related Concept Videos

Imaging Biological Samples with Optical Microscopy01:18

Imaging Biological Samples with Optical Microscopy

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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.
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Two basic types of preparation are used to visualize specimens with a light microscope: wet mounts and fixed specimens.
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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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Updated: Feb 26, 2026

High-resolution Episcopic Microscopy HREM - Simple and Robust Protocols for Processing and Visualizing Organic Materials
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High-resolution Episcopic Microscopy (HREM) - Simple and Robust Protocols for Processing and Visualizing Organic

Stefan H Geyer1, Barbara Maurer-Gesek1, Lukas F Reissig1

  • 1Division of Anatomy, Center for Anatomy and Cell Biology & MIC, Medical University of Vienna.

Journal of Visualized Experiments : Jove
|July 18, 2017
PubMed
Summary
This summary is machine-generated.

Simple protocols for high-resolution episcopic microscopy (HREM) generate digital volume data from organic materials. This method enables detailed 3D reconstruction and virtual sectioning of diverse specimens for advanced research.

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Area of Science:

  • Microscopy and Imaging Technologies
  • Biotechnology
  • Materials Science

Background:

  • High-resolution episcopic microscopy (HREM) offers advanced imaging capabilities for organic materials.
  • Generating digital volume data is crucial for 3D reconstruction and analysis.
  • Existing protocols may lack simplicity or tissue specificity.

Purpose of the Study:

  • To present simple, tissue-specific protocols for generating digital volume data using HREM.
  • To enable high-resolution 3D reconstruction of various organic specimens.
  • To facilitate virtual sectioning and model creation for research.

Main Methods:

  • Specimens embedded in methacrylate resin and sectioned using a microtome.
  • Digital images captured of block surfaces after each section using a video camera.
  • Image series processed for conversion to digital volume data with 3D visualization software.

Main Results:

  • Developed simple HREM protocols for diverse organic specimens, including embryos and human biopsy material.
  • Achieved typical numeric resolutions between 1 x 1 x 1 and 5 x 5 x 5 µm³.
  • Generated inherently aligned digital image series for immediate 3D data conversion.

Conclusions:

  • The presented HREM protocols are effective for generating high-quality digital volume data.
  • This method allows for detailed virtual sectioning and 3D model creation.
  • The protocols are applicable to a wide range of biological and material samples.