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Fluorescence and Phosphorescence: Instrumentation01:25

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Fluorometers and spectrofluorometers are two types of instruments used for measuring molecular fluorescence. These instruments differ in how they select excitation and emission wavelengths and the type of light sources they utilize. Fluorometers use absorption interference filters to choose excitation and emission wavelengths. The excitation source in a fluorometer is typically a low-pressure mercury vapor lamp that emits intense lines distributed throughout the ultraviolet and visible regions.
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Photoluminescence offers a wide range of applications due to its inherent sensitivity and selectivity. This technique allows for both direct and indirect analyses of the analyte. Direct quantitative analysis is possible when the analyte exhibits a favorable quantum yield for fluorescence or phosphorescence. However, an indirect analysis may be feasible if the analyte is not fluorescent or phosphorescent, or if the quantum yield is unfavorable. Indirect methods include reacting the analyte with...
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Development of Time-Resolved Luminescence Measurement Instruments for Biosensing and Bioimaging - An Overview.

Benjamin Sreenan1, Vala Kafil1, Donovan Wells1

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Time-resolved luminescence measurement (TRLM) uses long-lived probes for high-contrast biosensing. This review covers TRLM instruments, their development, and trends toward portable applications.

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Area of Science:

  • Biomedical Engineering
  • Analytical Chemistry
  • Optical Physics

Background:

  • Time-resolved luminescence measurement (TRLM) utilizes probes with decay times >100 nanoseconds for enhanced biosensing and bioimaging.
  • The pulse-delay-detect method in TRLM significantly improves signal-to-background ratios by minimizing light scattering and autofluorescence.

Purpose of the Study:

  • To review the recent development and applications of TRLM instruments for long-lived luminescence probes.
  • To explore instrument development trends towards point-of-care (POC) and in-field applications.
  • To analyze the impact of probe optical properties on TRLM instrument complexity, cost, compactness, and performance.

Main Methods:

  • Review of existing literature on TRLM instruments and long-lived luminescence probes.
  • Analysis of instrument design factors influenced by probe characteristics (e.g., decay time, emission spectrum).
  • Discussion of technological advancements in light sources, detectors, and electronics for TRLM.

Main Results:

  • A wide range of long-lived probes (organic dyes, lanthanide complexes, quantum dots, nanoparticles) are available for TRLM.
  • Various TRLM instruments have been developed, with emerging trends towards miniaturization and portability.
  • Probe properties critically influence instrument design, cost, and performance.

Conclusions:

  • TRLM offers a powerful approach for high-contrast bio-applications.
  • Further advancements in TRLM instrumentation are needed for low-cost, compact, and high-performance systems.
  • This review aims to foster a deeper understanding of TRLM instruments for broader adoption in biosensing and bioimaging.