Optical imaging in biomedical research: guidelines and practical insights

Preclinical imaging is crucial in biomedical research for non-invasive monitoring of various parameters in live animals over time, providing vital information while minimizing animal use. Optical imaging, the most commonly employed preclinical tool, offers ease of use, cost-effective instrumentation, and diverse applications. However, these advantages complicate the rational design, execution, and analysis of optical imaging experiments. This review highlights the specific conditions, strengths, and weaknesses of bioluminescence and fluorescence imaging. We discuss critical parameters essential for optimizing imaging settings to ensure accurate experimental outcomes. Additionally, we provide guidelines and address intrinsic limitations to assist scientists in making informed decisions and avoiding potential data shortcomings.