Enhancing precision in time-domain fluorescence lifetime imaging.
In biological applications of fluorescence lifetime imaging, low signals from samples can be a challenge, causing poor lifetime precision. We demonstrate how optimal signal gating (a method applied to the temporal dimension of a lifetime image) and novel total variation denoising models (a method applied to the spatial dimension of a lifetime image) can be used in time-domain fluorescence lifetime imaging microscopy (FLIM) to improve lifetime precision. In time-gated FLIM, notable fourfold precision improvements were observed in a low-light example. This approach can be employed to improve FLIM data while minimizing sample light exposure and increasing imaging speed.
University of Michigan, Department of Biomedical Engineering, Ann Arbor, Michigan 48109-2099, USA.
Ching-Wei
CW
Chang
Mary-Ann
MA
Mycek
D000465:Algorithms; D000818:Animals; D056808:Biostatistics; D003627:Data Interpretation, Statistical; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000706:statistics & numerical data; D008856:Microscopy, Fluorescence / Q000379:methods* / Q000706:statistics & numerical data; D057054:Molecular Imaging / Q000379:methods* / Q000706:statistics & numerical data; D055620:Optical Phenomena