Bioluminescence imaging: looking beyond the light.

Area of Science:

• Biomedical imaging
• Molecular biology
• Biophysics

Background:

• Bioluminescence imaging (BLI) is increasingly popular for in vivo molecular and cellular process visualization.
• Its sensitivity, ease of use, and in vitro to in vivo translatability contribute to its widespread adoption.
• Limited experience with BLI in living subjects has led to reported biases and unexpected readouts.

Purpose of the Study:

• To review biological and physical factors affecting bioluminescence imaging (BLI) accuracy.
• To provide researchers with knowledge to mitigate biases and improve BLI data interpretation.
• To highlight areas for future development in BLI techniques and compounds.

Main Methods:

• Literature review summarizing known influencing factors in bioluminescence imaging.
• Categorization of factors into biological (enzyme level) and physical (light propagation) effects.
• Analysis of reported challenges and their impact on BLI readout.

Main Results:

• Identified key biological factors influencing BLI signal generation at the enzyme level.
• Detailed physical factors affecting light propagation from the source to the detector.
• Highlighted the impact of these factors on the accuracy and interpretation of BLI data.

Conclusions:

• Understanding enzyme-level and light propagation factors is crucial for accurate bioluminescence imaging (BLI).
• Addressing these factors through improved strategies and compounds will enhance BLI's reliability in vivo.
• Future research should focus on mitigating identified biases for more precise molecular and cellular imaging.