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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.
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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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Conducting Multiple Imaging Modes with One Fluorescence Microscope
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Published on: October 28, 2018

Fluorescence interferometry: principles and applications in biology.

Alberto Bilenca1, Jing Cao, Max Colice

  • 1Harvard Medical School and Wellman Center for Photomedicine, Massachusetts General Hospital, 50 Blossom St., BAR 720, Boston, Massachusetts 02114, USA. abilenca@partners.org

Annals of the New York Academy of Sciences
|July 4, 2008
PubMed
Summary
This summary is machine-generated.

We introduce fluorescence interferometry to measure the phase information of fluorescent light waves. This new technique enables advanced nanoscale imaging and sensing in life sciences.

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Area of Science:

  • Optics
  • Biophysics
  • Nanotechnology

Background:

  • Fluorescence radiation is crucial for nanoscale measurements in life sciences.
  • Current methods primarily use fluorescence intensity, neglecting phase information.
  • Measuring fluorescence phase offers unexplored potential for advanced applications.

Purpose of the Study:

  • Introduce fluorescence interferometry for measuring fluorescence phase.
  • Explore its potential for novel imaging, sensing, ranging, and profiling methods.
  • Demonstrate experimental realizations and applications.

Main Methods:

  • Developed fluorescence interferometry, a form of optical low-coherence interferometry using fluorophores.
  • Proposed and tested two experimental setups to detect fluorescence interference patterns.
  • Compared capabilities and limitations with existing optical low-coherence interferometry schemes.

Main Results:

  • Achieved wide-field cross-sectional imaging with high resolution and large depth range.
  • Demonstrated quantitative profiling with nanometer-level precision.
  • Presented experimental evidence of fluorescence interferometry's practical applications.

Conclusions:

  • Fluorescence interferometry unlocks new possibilities for nanoscale measurements in life sciences.
  • It offers a powerful tool for advanced imaging, sensing, and profiling.
  • Future directions include fluorescence tomography and molecular interferometry.