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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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Confocal Fluorescence Microscopy01:16

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Confocal microscopy is an advanced microscopic technique. The prime advantage of the confocal microscope over other microscopy techniques is its ability to block the out-of-focus light from the illuminated samples using pinholes. It is widely used with fluorescence optics to obtain high-resolution, sharp contrast images. Unlike optical microscopes, confocal microscopes use a focused beam of light laser to scan the entire sample surface at different z-planes. These microscopes are, therefore,...
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Imaging Biological Samples with Optical Microscopy01:18

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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-Dimensional Microscopy in Microbiology01:29

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Two-dimensional (2D) microscopy encompasses a range of optical techniques that capture images within a single focal plane, offering detailed representations of microscopic structures. These techniques are essential in biological and medical research, enabling the visualization of cellular and subcellular structures with different levels of contrast and specificity.There are several major types of 2D microscopy, each with strengths and applications.Bright-Field MicroscopyBright-field microscopy...
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Phase Contrast and Differential Interference Contrast Microscopy01:26

Phase Contrast and Differential Interference Contrast Microscopy

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Phase-Contrast Microscopes
In-phase-contrast microscopes, interference between light directly passing through a cell and light refracted by cellular components is used to create high-contrast, high-resolution images without staining. It is the oldest and simplest type of microscope that creates an image by altering the wavelengths of light rays passing through the specimen. Altered wavelength paths are created using an annular stop in the condenser. The annular stop produces a hollow cone of...
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Electron Microscope Tomography and Single-particle Reconstruction01:07

Electron Microscope Tomography and Single-particle Reconstruction

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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.
Electron Tomography
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Related Experiment Video

Updated: Apr 21, 2026

Conducting Multiple Imaging Modes with One Fluorescence Microscope
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A high performance, cost-effective, open-source microscope for scanning two-photon microscopy that is modular and

David G Rosenegger1, Cam Ha T Tran1, Jeffery LeDue2

  • 1Department of Physiology and Pharmacology, Cumming School of Medicine, University of Calgary, Hotchkiss Brain Institute, Calgary, Alberta, Canada.

Plos One
|October 22, 2014
PubMed
Summary

Scientists can now build their own high-performance two-photon laser scanning microscopes. This open-source guide provides hardware, protocols, and 3D models to overcome cost and adaptability barriers for advanced biological imaging.

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Area of Science:

  • Neuroscience
  • Biophysics
  • Microscopy

Background:

  • Two-photon laser scanning microscopy is crucial for cellular and physiological studies.
  • High cost and limited adaptability of commercial systems hinder widespread adoption.
  • Need for accessible, high-performance imaging solutions for research laboratories.

Purpose of the Study:

  • To provide a detailed open-source hardware resource and protocol for building a modular two-photon microscope.
  • To enable researchers to construct adaptable, high-performance imaging systems.
  • To lower barriers for laboratories seeking advanced two-photon imaging capabilities.

Main Methods:

  • Designed an upright, modular two-photon laser scanning fluorescence microscope.
  • Utilized high-end components with off-the-shelf opto-mechanical parts.
  • Developed complete 3D computer models and assembly instructions for non-expert users.

Main Results:

  • Achieved imaging depths approaching 1 mm into mouse brain tissue.
  • Demonstrated a signal-to-noise ratio superior to tested commercial two-photon systems.
  • Provided a comprehensive guide including parts lists, assembly, testing, and troubleshooting.

Conclusions:

  • The open-source two-photon microscope design significantly reduces barriers to high-performance imaging.
  • Empowers more laboratories to conduct advanced in vitro and in vivo studies.
  • Facilitates progress in understanding cellular and physiological functions in living systems.