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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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A versatile total internal reflection photometric detection cell for flow analysis.

Peter S Ellis1, Brady S Gentle, Michael R Grace

  • 1Water Studies Centre, School of Chemistry, P.O. Box 23, Monash University, Clayton, Victoria 3800, Australia.

Talanta
|July 7, 2009
PubMed
Summary
This summary is machine-generated.

A novel total internal reflection (TIR) flow cell offers sensitive photometric measurements by minimizing schlieren effects and bubble trapping. This advanced flow cell achieved a lower limit of detection for reactive phosphate compared to traditional methods.

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Area of Science:

  • Analytical Chemistry
  • Optical Physics
  • Chemical Engineering

Background:

  • Photometric measurements in flow analysis require cells tolerant to optical distortions (schlieren effects) and free from gas bubble interference.
  • Existing flow cells, such as multi-reflection (MR) and Z-cells, have limitations in schlieren tolerance, hydrodynamic dispersion, bubble trapping, or dynamic range.
  • Liquid core waveguide cells offer total internal reflection but may not possess all the desired features for robust flow analysis.

Purpose of the Study:

  • To describe a novel total internal reflection (TIR) flow-through cell designed for sensitive and reliable photometric measurements.
  • To evaluate the performance of the TIR flow cell in terms of schlieren tolerance, hydrodynamic dispersion, bubble trapping, and detection limits.
  • To compare the efficacy of the TIR flow cell against established MR and Z-cells for a specific analytical determination.

Main Methods:

  • Light introduction via optical fiber into a quartz capillary at a specific incident angle (ca. 53 degrees) for total internal reflection.
  • Propagation of light through the aqueous liquid core via successive reflections.
  • Detection using a second optical fiber and a charge-coupled device detector coupled via an optical coupling medium.
  • Application in a flow injection system for the determination of reactive phosphate.

Main Results:

  • The TIR flow cell demonstrated high tolerance to schlieren effects and minimal hydrodynamic dispersion and bubble trapping.
  • A limit of detection of 2.0 microg PL(-1) for reactive phosphate was achieved using the TIR cell.
  • This limit of detection was significantly lower than those obtained with MR (3.8 microg PL(-1)) and Z-cells (4.9 microg PL(-1)) under identical conditions.

Conclusions:

  • The developed TIR flow cell integrates the advantages of liquid core waveguides, MR cells, and Z-cells, offering superior performance for photometric measurements.
  • Its robustness against schlieren effects and bubble entrapment, combined with a high signal-to-noise ratio and wide dynamic range, makes it suitable for analyzing samples with diverse refractive indices.
  • The TIR flow cell represents a significant advancement for sensitive and reliable quantitative analysis in flow systems.