Open AccessEliminating deformations in fluorescence emission difference microscopy
Author(s) -
Shangting You,
Cuifang Kuang,
Zihao Rong,
Xu Liu
Publication year - 2014
Publication title -
optics express
Language(s) - Uncategorized
Resource type - Journals
SCImago Journal Rank - 1.394
H-Index - 271
ISSN - 1094-4087
DOI - 10.1364/oe.22.026375
Subject(s) - optics , microscopy , materials science , resolution (logic) , fluorescence lifetime imaging microscopy , fluorescence , confocal , confocal microscopy , light sheet fluorescence microscopy , fluorescence microscope , image resolution , microscope , background subtraction , image subtraction , image processing , physics , pixel , image (mathematics) , computer science , binary image , artificial intelligence
We propose a method for eliminating the deformations in fluorescence emission difference microscopy (FED). Due to excessive subtraction, negative values are inevitable in the original FED method, giving rise to deformations. We propose modulating the beam to generate an extended solid focal spot and a hollow focal spot. Negative image values can be avoided by using these two types of excitation spots in FED imaging. Hence, deformations are eliminated, and the signal-to-noise ratio is improved. In deformation-free imaging, the resolution is higher than that of confocal imaging by 32%. Compared to standard FED imaging with the same level of deformations, our method provides superior resolution.