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Related Concept Videos

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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Overview of Exosomes01:36

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Exosomes are stable, lipid bilayer-enclosed vesicles capable of crossing biological barriers. They can carry a wide range of molecules required for intercellular communication. Once exosomes are released from the cell where they originated, they enter a recipient cell through various pathways such as fusion, receptor-mediated endocytosis, macropinocytosis, and phagocytosis.
Stahl et al. discovered exosomes in 1983, but the exosomes were initially considered waste products released from the...
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Related Experiment Video

Updated: Nov 3, 2025

Using Nanoplasmon-Enhanced Scattering and Low-Magnification Microscope Imaging to Quantify Tumor-Derived Exosomes
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Detection of Exosomes Using Total Internal Reflected Imaging Ellipsometry.

Haoyu Liu1,2, Wei Liu1, Gang Jin1,2

  • 1NML, Institute of Mechanics, Chinese Academy of Sciences, 15 Bei-Si-Huan West Road, Beijing 100190, China.

Biosensors
|June 2, 2021
PubMed
Summary
This summary is machine-generated.

Researchers developed a new biosensing surface for detecting exosomes, tiny vesicles involved in cell communication and disease. This method uses total internal reflected imaging ellipsometry (TIRIE) and identifies CD9 as a more suitable exosome surface marker than CD63.

Keywords:
CD63CD9TIRIEexosomesinteraction kineticssurface protein

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Sample Preparation and Imaging of Exosomes by Transmission Electron Microscopy
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Area of Science:

  • Biotechnology
  • Nanotechnology
  • Biochemistry

Background:

  • Exosomes are critical nanovesicles in biological fluids, mediating intercellular communication and playing roles in physiology and pathology.
  • Their potential as disease biomarkers necessitates sensitive and specific detection methods.
  • Existing methods often lack the required precision for exosome analysis.

Purpose of the Study:

  • To develop an efficient biosensing surface for capturing and detecting exosomes.
  • To compare the suitability of two exosome surface proteins, CD9 and CD63, as detection markers.
  • To evaluate the performance of total internal reflected imaging ellipsometry (TIRIE) for real-time exosome analysis.

Main Methods:

  • Development of a biosensing surface by immobilizing Protein G and antibodies on a golden substrate.
  • Utilizing total internal reflected imaging ellipsometry (TIRIE) for real-time monitoring of bio-interactions.
  • Analysis of exosome binding kinetics using pseudo-first-order interaction kinetics to determine dissociation constants (KD).

Main Results:

  • The developed biosensing surface effectively captured exosomes, with an average adhered exosome distance of 44 nm ± 0.5 nm.
  • Antibody against CD9 exhibited a lower dissociation constant (KD) compared to anti-CD63, indicating higher binding affinity.
  • The limit of detection (LOD) for exosomes using TIRIE was determined to be 0.4 μg/mL.

Conclusions:

  • A novel biosensing surface based on TIRIE has been successfully developed for exosome detection.
  • CD9 is identified as a more suitable surface marker for exosome detection compared to CD63 using this method.
  • The proposed TIRIE-based approach offers a convenient and efficient platform for exosome analysis, with potential applications in biomarker discovery.