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

All Science Journal Classification (ASJC) codes

  • Atomic and Molecular Physics, and Optics
  • Instrumentation

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