title: Masked illumination scheme for a galvanometer scanning high-speed confocal fluorescence microscope.
abstract: High-speed beam scanning and data acquisition in a laser scanning confocal microscope system are normally implemented with a resonant galvanometer scanner and a frame grabber. However, the nonlinear scanning speed of a resonant galvanometer can generate nonuniform photobleaching in a fluorescence sample as well as image distortion near the edges of a galvanometer scanned fluorescence image. Besides, incompatibility of signal format between a frame grabber and a point detector can lead to digitization error during data acquisition. In this article, we introduce a masked illumination scheme which can effectively decrease drawbacks in fluorescence images taken by a laser scanning confocal microscope with a resonant galvanometer and a frame grabber. We have demonstrated that the difference of photobleaching between the center and the edge of a fluorescence image can be reduced from 26 to 5% in our confocal laser scanning microscope with a square illumination mask. Another advantage of our masked illumination scheme is that the zero level or the lowest input level of an analog signal in a frame grabber can be accurately set by the dark area of a mask in our masked illumination scheme. We have experimentally demonstrated the advantages of our masked illumination method in detail.
authors.affiliation: School of Information and Mechatronics, Gwangju Institute of Science and Technology, Gwangju, Republic of Korea.
authors.forename: Dong Uk
authors.initials: DU
authors.lastname: Kim
authors.forename: Sucbei
authors.initials: S
authors.lastname: Moon
authors.forename: Hoseong
authors.initials: H
authors.lastname: Song
authors.forename: Hyuk-Sang
authors.initials: HS
authors.lastname: Kwon
authors.forename: Dug Young
authors.initials: DY
authors.lastname: Kim
mesh_terms: D001773:Blood Cells / Q000648:ultrastructure; D006801:Humans; D008029:Lighting / Q000379:methods*; D018613:Microscopy, Confocal / Q000379:methods*