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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,...
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.
Phase Contrast and Differential Interference Contrast Microscopy01:26

Phase Contrast and Differential Interference Contrast Microscopy

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...
Imaging Biological Samples with Optical Microscopy01:18

Imaging Biological Samples with Optical Microscopy

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.
In optical microscopy, the specimen to be viewed is placed on a glass slide and clipped on the stage...

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Related Experiment Video

Updated: Jun 21, 2026

Simultaneous Multicolor Imaging of Biological Structures with Fluorescence Photoactivation Localization Microscopy
12:51

Simultaneous Multicolor Imaging of Biological Structures with Fluorescence Photoactivation Localization Microscopy

Published on: December 9, 2013

Focal modulation microscopy.

Nanguang Chen1, Chee-Howe Wong, Colin J R Sheppard

  • 1Division of Bioengineering, National University of Singapore, Singapore. biecng@nus.edu.sg

Optics Express
|July 8, 2009
PubMed
Summary
This summary is machine-generated.

We developed a new light microscopy technique for high-resolution molecular imaging in thick tissues. This method effectively images deep within scattering tissues by suppressing background noise, achieving a depth of 600 microns.

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Area of Science:

  • Biomedical Optics
  • Molecular Imaging
  • Microscopy

Background:

  • Imaging deep within thick biological tissues is challenging due to light scattering and background fluorescence.
  • Conventional microscopy techniques struggle to achieve high resolution and effective optical sectioning in scattering media.

Purpose of the Study:

  • To introduce a novel light microscopy method for high-resolution molecular imaging of thick biological tissues.
  • To demonstrate effective optical sectioning and diffraction-limited spatial resolution deep within scattering tissues.

Main Methods:

  • Utilized one-photon excited fluorescence microscopy.
  • Employed focal modulation, a technique to suppress background fluorescence excited by scattered light.
  • Validated the method on animal tissue samples.

Main Results:

  • Achieved effective optical sectioning and diffraction-limited spatial resolution.
  • Successfully imaged deep within multiple-scattering biological tissues.
  • Demonstrated an imaging depth of approximately 600 microns.

Conclusions:

  • The developed light microscopy method enables high-resolution molecular imaging in thick tissues.
  • Focal modulation is effective in overcoming scattering and background noise for deep tissue imaging.
  • This technique offers a promising tool for studying biological processes in intact tissues.