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Related Concept Videos

Confocal Fluorescence Microscopy01:16

Confocal Fluorescence Microscopy

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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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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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Total Internal Reflection Fluorescence Microscopy01:05

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Total internal reflection fluorescence microscopy or TIRF is an advanced microscopic technique used to visualize fluorophores in samples close to a solid surface with a higher refractive index, such as a glass coverslip. TIRF only allows fluorophores in proximity to the solid surface to be excited. When light from a medium with a lower refractive index (such as air) hits the glass coverslip at a critical angle, the light undergoes total internal reflection stead of passing through the glass.
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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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Cells with similar structure and function are grouped into tissues. A group of tissues with a specialized function is called an organ. There are four main types of tissue in vertebrates: epithelial, connective, muscle, and nervous.
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Tissues are a group of cells that share a common embryonic origin. Microscopic observation reveals that the cells in a tissue share morphological features and are arranged in an orderly pattern to perform specific functions. From an evolutionary perspective, tissues appear in more complex organisms. Although there are many types of cells in the human body, they are organized into four broad categories of tissues: epithelial, connective, muscle, and nervous. Each of these categories is...
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Related Experiment Video

Updated: Feb 15, 2026

Deep-Tissue Three-Photon Fluorescence Microscopy in Intact Mouse and Zebrafish Brain
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Deep Tissue Imaging with Multiphoton Fluorescence Microscopy.

David R Miller1, Jeremy W Jarrett1, Ahmed M Hassan1

  • 1Department of Biomedical Engineering, The University of Texas at Austin, 107 W. Dean Keeton C0800, Austin, TX 78712, USA.

Current Opinion in Biomedical Engineering
|January 17, 2018
PubMed
Summary
This summary is machine-generated.

Multiphoton microscopy enables deep-tissue imaging by optimizing excitation wavelengths (1,300 nm and 1,700 nm) and selecting appropriate fluorophores and laser sources for neuroscience applications.

Keywords:
Fluorophores for multiphoton microscopyLaser sources for multiphoton microscopyMultiphoton Microscopy Fluorescence ImagingScattering and absorption in brain tissue

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Area of Science:

  • Biomedical Optics
  • Microscopy Techniques
  • Neuroscience Imaging

Background:

  • Deep-tissue imaging is crucial for understanding biological structures and functions.
  • Light scattering and absorption in biological tissues limit imaging depth and resolution.
  • Multiphoton microscopy offers advantages for deep-tissue visualization compared to traditional methods.

Purpose of the Study:

  • To review advancements in deep-tissue imaging using multiphoton microscopy.
  • To identify optimal excitation wavelengths and suitable fluorophores for enhanced imaging.
  • To discuss the utility of multiphoton microscopy in neuroscience research.

Main Methods:

  • Analysis of optical properties of biological samples influencing light propagation.
  • Identification of ideal excitation wavelengths (1,300 nm and 1,700 nm) based on scattering and absorption characteristics.
  • Review of available fluorophores and ultrafast laser sources for multiphoton excitation.

Main Results:

  • 1,300 nm and 1,700 nm wavelengths are identified as optimal for deep-tissue multiphoton imaging.
  • A summary of current fluorophore availability and ultrafast laser technologies is provided.
  • The review highlights the potential of multiphoton microscopy for neuroscience.

Conclusions:

  • Multiphoton microscopy, utilizing specific wavelengths and advanced light sources, significantly improves deep-tissue imaging capabilities.
  • The selection of appropriate fluorophores is critical for successful multiphoton imaging.
  • This technique holds substantial promise for advancing neuroscience research and understanding brain function.