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Related Concept Videos

Confocal Fluorescence Microscopy01:16

Confocal Fluorescence Microscopy

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,...
Immunofluorescence Microscopy01:12

Immunofluorescence Microscopy

A fluorescence microscope uses fluorescent chromophores called fluorochromes, which can absorb energy from a light source and then emit this energy as visible light. Fluorochromes include naturally fluorescent substances (such as chlorophylls) and fluorescent stains that are added to the specimen to create contrast. Dyes such as Texas red and FITC are examples of fluorochromes. Other examples include the nucleic acid dyes 4’,6’-diamidino-2-phenylindole (DAPI), and acridine orange.
The...
Two-Dimensional Microscopy in Microbiology01:29

Two-Dimensional Microscopy in Microbiology

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...
Super-resolution Fluorescence Microscopy01:37

Super-resolution Fluorescence Microscopy

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 developed.
Three-Dimensional Microscopy in Microbiology01:28

Three-Dimensional Microscopy in Microbiology

Three-dimensional imaging techniques are essential in cell biology, allowing researchers to visualize intricate cellular structures with high resolution. Two prominent methods, Differential Interference Contrast Microscopy (DIC) and Confocal Scanning Laser Microscopy (CSLM), provide distinct advantages for imaging live and thick specimens, respectively.Differential Interference Contrast MicroscopyDIC microscopy enhances contrast in transparent, unstained samples by converting phase...
Overview of Microscopy Techniques01:22

Overview 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...

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Updated: Jun 17, 2026

Conducting Multiple Imaging Modes with One Fluorescence Microscope
08:32

Conducting Multiple Imaging Modes with One Fluorescence Microscope

Published on: October 28, 2018

Fluorescence microscopy: a concise guide to current imaging methods.

Christian A Combs1

  • 1NHLBI Light Microscopy Facility, NIH, Bethesda, Maryland.

Current Protocols in Neuroscience
|January 13, 2010
PubMed
Summary
This summary is machine-generated.

Choosing the right fluorescence microscopy technique can be challenging. This guide explains advanced imaging methods like structured light, confocal, TIRF, two-photon, and STED microscopy for cell and neurobiology research.

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Imaging the Human Immunological Synapse
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Fluorescence Imaging with One-nanometer Accuracy (FIONA)

Published on: September 26, 2014

Related Experiment Videos

Last Updated: Jun 17, 2026

Conducting Multiple Imaging Modes with One Fluorescence Microscope
08:32

Conducting Multiple Imaging Modes with One Fluorescence Microscope

Published on: October 28, 2018

Imaging the Human Immunological Synapse
09:37

Imaging the Human Immunological Synapse

Published on: December 26, 2019

Fluorescence Imaging with One-nanometer Accuracy (FIONA)
11:56

Fluorescence Imaging with One-nanometer Accuracy (FIONA)

Published on: September 26, 2014

Area of Science:

  • Life Sciences
  • Biophysics
  • Microscopy

Background:

  • Fluorescence microscopy offers advanced imaging capabilities for cell and neurobiology.
  • Rapid technological advancements present challenges for non-expert users in selecting appropriate techniques.
  • Understanding imaging physics and balancing properties is crucial for effective biological research.

Purpose of the Study:

  • To provide concise descriptions of various commercially available fluorescence microscopy techniques.
  • To offer a comparative guide for selecting the optimal imaging method based on biological questions.
  • To aid non-expert microscopists in making informed decisions for their research.

Main Methods:

  • Descriptions of structured light microscopy.
  • Explanations of confocal microscopy.
  • Details on total internal reflection fluorescence (TIRF) microscopy.
  • Overview of two-photon microscopy.
  • Introduction to stimulated emission depletion (STED) microscopy.

Main Results:

  • Concise descriptions of key fluorescence microscopy techniques are presented.
  • A tabular guide facilitates the comparison of imaging properties.
  • Techniques covered include structured light, confocal, TIRF, two-photon, and STED microscopy.

Conclusions:

  • Selecting the appropriate fluorescence microscopy technique requires understanding imaging physics and balancing competing properties.
  • This unit serves as a practical resource for researchers, particularly non-experts.
  • Informed technique selection enhances the ability to address specific biological questions effectively.