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Related Concept Videos

Total Internal Reflection Fluorescence Microscopy01:05

Total Internal Reflection Fluorescence Microscopy

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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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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Computed Tomography-guided Time-domain Diffuse Fluorescence Tomography in Small Animals for Localization of Cancer Biomarkers
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Fluorescence tomographic microscopy by wavefront detection.

Ignacio Iglesias1

  • 1Departamento de Física, Universidad de Murcia, Campus de Espinardo (CIOyN bldg.), E-30100 Murcia, Spain. iic@um.es

Optics Letters
|April 6, 2010
PubMed
Summary
This summary is machine-generated.

Researchers developed a novel real-time method for direct depth information acquisition in scanning fluorescence microscopy. This technique utilizes axial excitation and detects emitted light

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Area of Science:

  • Optics and Photonics
  • Biomedical Imaging
  • Microscopy Techniques

Background:

  • Scanning fluorescence microscopy is a crucial technique for biological imaging.
  • Acquiring real-time depth information in such microscopy remains a challenge.
  • Existing methods often require complex setups or are limited in speed.

Purpose of the Study:

  • To introduce a new, direct method for real-time depth information retrieval in scanning fluorescence microscopy.
  • To demonstrate the feasibility and performance of this novel approach.

Main Methods:

  • The method employs axial excitation of the sample.
  • It relies on detecting both the phase and intensity of the emitted fluorescence light.
  • An axicon lens was used to generate a Bessel beam for excitation, and a microlens array served as the phase detector.

Main Results:

  • Successful real-time acquisition of depth information was achieved.
  • The use of Bessel beams and microlens arrays proved effective for phase detection.
  • The presented results validate the proposed method's capability.

Conclusions:

  • A new method for real-time direct depth information in scanning fluorescence microscopy has been successfully developed.
  • The technique offers a promising advancement for 3D biological imaging.
  • Further applications in advanced microscopy are anticipated.