Luminescence lifetime imaging of three-dimensional biological objects

Area of Science:

• Biophotonics and advanced optical imaging techniques.
• Multiscale biological and molecular analysis.

Background:

• Bridging knowledge gaps between cellular, tissue, and organism scales is a key challenge in biology.
• Optical analytical tools enable multiparameter quantitative imaging of biological systems.
• Luminescence lifetime imaging modalities offer significant insights.

Purpose of the Study:

• To provide an overview of luminescence lifetime imaging techniques.
• To summarize key biological applications of luminescence lifetime imaging.
• To highlight the utility of these methods in bridging scale-dependent knowledge gaps.

Main Methods:

• Utilizing luminescence lifetime imaging, including fluorescence lifetime imaging (FLI) and phosphorescence lifetime imaging microscopy (PLIM).
• Employing Förster resonance energy transfer (FRET) assays in synergy with lifetime imaging.
• Quantitative imaging of live and fixed cells, 3D systems, tissues, and organisms.

Main Results:

• Luminescence lifetime imaging provides molecular insights into protein-protein interactions, cellular metabolism, oxygen dynamics, and ion concentrations.
• These techniques enable functional and molecular analysis across multiple scales (microscopic to macroscopic).
• 3D spatially resolved and longitudinal measurements are achievable.

Conclusions:

• Luminescence lifetime imaging offers a unique window into the physiological and structural environment of biological systems.
• It enables a new level of functional and molecular analysis from cells to whole organisms.
• These advanced imaging methods are crucial for comprehensive biological studies across scales.