Related Experiment Video
Updated: Mar 17, 2026
14:20
Absolute Quantum Yield Measurement of Powder Samples
Published on: May 12, 2012
28.8K
Quantum Yield Determination Based on Photon Number Measurement, Protocols for Firefly Bioluminescence Reactions
1Quantum Optical Measurement Group, National Metrology Institute of Japan, National Institute of Advanced Industrial Science and Technology, 1-1-1 Umezono, Tsukuba, Ibaraki, 563-8563, Japan. niwa-k@aist.go.jp.
Methods in Molecular Biology (Clifton, N.J.)
|July 19, 2016
Summary
This study details methods for measuring photon counts to determine quantum yield (QY) in bioluminescence reactions, using firefly luminescence as an example. Accurate QY measurement is crucial for understanding light intensity in luminescence assays.
Area of Science:
- Photochemistry
- Biochemistry
- Radiometry
Background:
- Quantum yield (QY) quantifies photon production efficiency in luminescence reactions.
- Accurate QY is essential for characterizing light intensity from bio/chemiluminescence.
- Standardized measurement methods are needed for reliable QY determination.
Purpose of the Study:
- To describe methods for measuring photon counts to determine quantum yield (QY).
- To align QY measurement techniques with national radiometry standards.
- To provide an experimental example using firefly bioluminescence.
Main Methods:
- Photon counting techniques based on national radiometry standards.
- Application of these methods to firefly bioluminescence reactions.
- Calibration and validation of the measurement system.
Main Results:
- Demonstrated a method for accurate photon number determination.
- Successfully applied the method to measure QY in firefly bioluminescence.
- Established a protocol for QY measurement aligned with radiometry standards.
Conclusions:
- The described methods enable precise QY determination for luminescence reactions.
- Standardized measurements are vital for comparing luminescence intensities across studies.
- Firefly bioluminescence serves as a reliable model for validating these QY measurement techniques.
Related Concept Videos
01:14Photoluminescence: Applications
1.2K
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...
1.2K
01:25Fluorescence and Phosphorescence: Instrumentation
1.8K
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.
1.8K