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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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Updated: Jul 7, 2026

Fluorescence detection methods for microfluidic droplet platforms
14:16

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Published on: December 10, 2011

Exciplex fluorescence imaging for liquid mixing studies.

K T Chojnacki, D A Feikema

    Applied Optics
    |February 15, 2008
    PubMed
    Summary

    This study introduces an excited-state complex (exciplex) fluorescence technique for visualizing liquid diffusion. The method offers high-resolution imaging, providing insights into mixing dynamics and diffusion scales.

    Area of Science:

    • Fluid dynamics
    • Chemical physics
    • Optical diagnostics

    Background:

    • Understanding liquid mixing is crucial in various scientific and industrial applications.
    • Visualizing diffusion layers in real-time presents significant challenges.
    • Current methods may lack the spatial or temporal resolution needed to capture fine-scale mixing phenomena.

    Purpose of the Study:

    • To develop and demonstrate a novel diagnostic technique for visualizing diffusion layers during liquid mixing.
    • To provide high-spatial-resolution instantaneous and time-sequenced images of mixing processes.
    • To obtain time-resolved data on the duration and scales of diffusion events.

    Main Methods:

    • Utilizing excited-state complex (exciplex) fluorescence as the core diagnostic principle.

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    Fluorescence detection methods for microfluidic droplet platforms
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    Microfluidic Mixers for Studying Protein Folding

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  • Employing high-spatial-resolution imaging to capture instantaneous and time-sequenced visualizations.
  • Applying time-resolved imaging to analyze the temporal dynamics of mixing.

    • Main Results:

    • Successful visualization of diffusion layers formed between mixing liquids.
    • High-resolution images of single drops and jets mixing within a liquid pool were obtained.
    • Time-resolved exciplex fluorescence images provided data on mixing event duration and diffusion length/time scales.

    Conclusions:

    • The developed exciplex fluorescence technique is effective for visualizing liquid diffusion layers.
    • The technique offers potential for new insights into the physics of liquid mixing processes.
    • This method can provide valuable information on the temporal and spatial aspects of diffusion.