Fluorescence lifetime images and correlation spectra obtained by multidimensional time-correlated single photon...

Area of Science:

• Biophysics
• Spectroscopy
• Microscopy

Background:

• Multidimensional time-correlated single photon counting (TCSPC) is a sophisticated fluorescence detection technique.
• It involves exciting samples with a high-repetition rate laser and detecting single photons across multiple channels.

Purpose of the Study:

• To demonstrate the versatility of multidimensional TCSPC.
• To show its application in both fluorescence lifetime analysis and fluorescence correlation spectroscopy (FCS).

Main Methods:

• Utilizing a confocal or two-photon laser scanning microscope.
• Employing a pulsed laser system for sample excitation.
• Characterizing detected photons by arrival time, channel, and spatial/temporal coordinates.

Main Results:

• Multidimensional TCSPC offers multiwavelength capabilities and high counting efficiency.
• The technique can resolve complex, multiexponential fluorescence decay functions.
• The same hardware successfully performs both precision fluorescence decay analysis and FCS.

Conclusions:

• Multidimensional TCSPC is a powerful and adaptable technique for advanced fluorescence studies.
• It provides a unified platform for fluorescence lifetime and correlation spectroscopy.
• This approach enhances the analytical capabilities in biophysical and materials science research.