Fluorescence Lifetime Imaging Techniques-A Review on Principles, Applications and Clinical Relevance

Area of Science:

• Microscopy and Imaging Technologies
• Biophysics
• Molecular Biology

Background:

• Fluorescence lifetime is a direct indicator of molecular environment interactions.
• Fluorescence-lifetime imaging (FLIM) offers molecular imaging capabilities beyond traditional contrast techniques.
• Understanding fluorescence and phosphorescence lifetime is crucial for advanced microscopy.

Purpose of the Study:

• To provide an overview of commonly used FLIM techniques.
• To discuss the capabilities and applications of various FLIM methods.
• To highlight the significance of FLIM in biological systems.

Main Methods:

• Classification of FLIM techniques into time-domain and frequency-domain methods.
• Categorization into analogue and photon counting techniques.
• Distinction between scanning and wide-field imaging approaches.

Main Results:

• Detailed description of the features and peculiarities of different FLIM techniques.
• Emphasis on Time-Correlated Single Photon Counting (TCSPC) FLIM.
• Identification of unique capabilities of TCSPC FLIM for biological applications.

Conclusions:

• FLIM is a powerful molecular imaging technique.
• Various FLIM techniques offer distinct advantages for different applications.
• TCSPC FLIM presents unique capabilities for studying biological systems.