Masked illumination scheme for a galvanometer scanning high-speed confocal fluorescence microscope

Dong Uk Kim, Sucbei Moon, Hoseong Song, Hyuk Sang Kwon, Dug Young Kim

Research output: Contribution to journalArticle

4 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)455-462
Number of pages8
JournalScanning
Volume33
Issue number6
DOIs
Publication statusPublished - 2011 Nov 1

Fingerprint

Galvanometers
galvanometers
Microscopes
Lighting
Fluorescence
illumination
microscopes
high speed
Scanning
fluorescence
scanning
Photobleaching
data acquisition
Lasers
Masks
Data acquisition
masks
lasers
incompatibility
Analog to digital conversion

All Science Journal Classification (ASJC) codes

  • Atomic and Molecular Physics, and Optics
  • Instrumentation

Cite this

Kim, Dong Uk ; Moon, Sucbei ; Song, Hoseong ; Kwon, Hyuk Sang ; Kim, Dug Young. / Masked illumination scheme for a galvanometer scanning high-speed confocal fluorescence microscope. In: Scanning. 2011 ; Vol. 33, No. 6. pp. 455-462.
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Masked illumination scheme for a galvanometer scanning high-speed confocal fluorescence microscope. / Kim, Dong Uk; Moon, Sucbei; Song, Hoseong; Kwon, Hyuk Sang; Kim, Dug Young.

In: Scanning, Vol. 33, No. 6, 01.11.2011, p. 455-462.

Research output: Contribution to journalArticle

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