Fluorescence lifetime imaging using multi-dimension time-correlated single photon counting method

2010 ◽  
Author(s):  
Cuixia Sheng ◽  
Heijing Tang
Keyword(s):  
Fluorescence Lifetime ◽  
Single Photon ◽  
Photon Counting ◽  
Fluorescence Lifetime Imaging ◽  
Counting Method ◽  
Single Photon Counting ◽  
Lifetime Imaging ◽  
Photon Counting Method

Signal Processing Technique of Fluorescence Lifetime Imaging Using Four-Dimension Time Correlated Single Photon Counting Method

2011 ◽  
Vol 216 ◽  
pp. 350-354
Author(s):  
Cui Xia Sheng ◽  
He Jing Tang ◽  
Hong Yan Jia ◽  
Shu Lian Yang
Keyword(s):  
Fluorescence Lifetime ◽  
Single Photon ◽  
Photon Counting ◽  
Fluorescence Lifetime Imaging ◽  
Counting Method ◽  
Single Photon Counting ◽  
Lifetime Imaging ◽  
Scanning Area ◽  
Multi Wavelength ◽  
Photon Counting Method

Measurement of fluorescence lifetime plays an important role in fluorescence lifetime imaging technology. If the Time correlated single photon counting method is used to process the signal of fluorescence lifetime images, it can make the real time imaging which have the virtue of high timing resolve and high signal to noise ratio. Multi-wavelength channels are used to record the lifetime simultaneously in four-dimension TCSPC system design. Four-dimension TCSPC imaging can be used to record the photon density over time, wavelength and coordinates of the scanning area. Analysis shows that the counting efficiency and the amount of information in the data can be increased by recording the fluorescence in several wavelength channels simultaneously. Multi-wavelength imaging was also successfully used to separate different fluorephores in stead-state images.

scholarly journals Optical Estimation of Absolute Membrane Potential Using One- and Two-Photon Fluorescence Lifetime Imaging Microscopy

2021 ◽  
Author(s):  
Julia R. Lazzari-Dean ◽  
Evan W. Miller
Keyword(s):  
Membrane Potential ◽  
Fluorescence Lifetime ◽  
Single Photon ◽  
Photon Counting ◽  
Fluorescence Lifetime Imaging ◽  
Single Photon Counting ◽  
Two Photon ◽  
Lifetime Imaging ◽  
Wide Range ◽  
Relative Measurements

AbstractBackgroundMembrane potential (Vmem) exerts physiological influence across a wide range of time and space scales. To study Vmem in these diverse contexts, it is essential to accurately record absolute values of Vmem, rather than solely relative measurements.Materials & MethodsWe use fluorescence lifetime imaging of a small molecule voltage sensitive dye (VF2.1.Cl) to estimate mV values of absolute membrane potential.ResultsWe test the consistency of VF2.1.Cl lifetime measurements performed on different single photon counting instruments and find that they are in striking agreement (differences of <0.5 ps/mV in the slope and <50 ps in the y-intercept). We also demonstrate that VF2.1.Cl lifetime reports absolute Vmem under two-photon (2P) illumination with better than 20 mV of Vmem resolution, a nearly 10-fold improvement over other lifetime-based methods.ConclusionsWe demonstrate that VF-FLIM is a robust and portable metric for Vmem across imaging platforms and under both one-photon and two-photon illumination. This work is a critical foundation for application of VF-FLIM to record absolute membrane potential signals in thick tissue.

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