Open AccessDescription of deep saturated excitation multiphoton microscopy for super-resolution imaging
Author(s) -
Genevieve D. Vigil,
Yide Zhang,
Aamir A. Khan,
Scott S. Howard
Publication year - 2017
Publication title -
journal of the optical society of america. a, optics, image science, and vision./journal of the optical society of america. a, online
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.803
H-Index - 158
eISSN - 1520-8532
pISSN - 1084-7529
DOI - 10.1364/josaa.34.001217
Subject(s) - microscopy , excitation , resolution (logic) , optics , diffraction , point spread function , image resolution , microscope , materials science , image quality , superresolution , temporal resolution , scattering , computer science , physics , image (mathematics) , artificial intelligence , quantum mechanics
Here we recount the standard two-level model that describes saturated excitation (SAX) in multiphoton microscopy (MPM), a new technique for super-resolution fluorescence microscopy in scattering tissue, which requires no special chemistry and only simple modifications to a commercial MPM microscope. We use the model to study conditions required for improvements in MPM SAX resolution and experimental implementation strategies. Simulation results find zeros, or nodes, in the frequency response, which generate highly irregular point-spread functions (PSFs), such as rings and ripples, that contain spatial frequency content >3× larger than allowed by diffraction. These PSFs are a direct result of zeros in the frequency response of saturated fluorophores under specific excitation conditions. The impact on image quality is discussed using simulations of targets imaged with SAX PSFs. Further, we explore engineering sets of irregular PSFs by varying the excitation power and reconstructing super-resolution images from the set of captured images.