Open AccessOptimizing depth-of-field extension in optical sectioning microscopy techniques using a fast focus-tunable lens
Author(s) -
Kuan-Lun Hsu,
Kuan-Yu Li,
Yen-Yin Lin,
AnnShyn Chiang,
ShiWei Chu
Publication year - 2017
Publication title -
optics express
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.394
H-Index - 271
ISSN - 1094-4087
DOI - 10.1364/oe.25.016783
Subject(s) - optics , depth of field , optical sectioning , lens (geology) , microscope , focus (optics) , depth of focus (tectonics) , holography , microscopy , axial symmetry , materials science , micrometer , confocal , computer science , physics , paleontology , subduction , quantum mechanics , biology , tectonics
Volume imaging based on a fast focus-tunable lens (FTL) allows three-dimensional (3D) observation within milliseconds by extending the depth-of-field (DOF) with sub-micrometer transverse resolution on optical sectioning microscopes. However, the previously published DOF extensions were neither axially uniform nor fit with theoretical prediction. In this work, complete theoretical treatments of focus extension with confocal and various multiphoton microscopes are established to correctly explain the previous results. Moreover, by correctly placing the FTL and properly adjusting incident beam diameter, a uniform DOF is achieved in which the actual extension nicely agrees with the theory. Our work not only provides a theoretical platform for volumetric imaging with FTL but also demonstrates the optimized imaging condition.