Fast and adaptive auto-focusing algorithm for microscopic cell observation

Takeshi Obara, Yasunobu Igarashi, Koichi Hashimoto

研究成果: Conference contribution

13 被引用数 (Scopus)

抄録

In biological and medical sciences, optical microscopes are widely used. By using the microscope, we can observe cellular dynamics including intracellular ions and molecules tagged with fluorescent dyes at a high magnification. However, a freely motile cell easily escapes from a field of view and a focal plane of the typical microscope. Therefore, we propose a novel auto-focusing (AF) algorithm and develop an AF and tracking microscope. XYZ positions of a microscopic stage are visual feedback controlled to focus and track the cell automatically. A bright-field image is used to estimate a cellular position. XY centroids are used to estimate XY positions of the tracked cell. To estimate a Z position, we use a diffraction pattern around the cell membrane. This estimation method is so-called Depth from Diffraction (DFDi). However, this method is not robust for individual differences between cells because the diffraction pattern depends on each cellular shape, reflective index and transparency. Therefore, in this study, we propose a real-time correction of DFDi by using 2D Laplacian of a center region of the cell as a goodness of the focus. To evaluate the performance of our developed algorithm and microscope, we auto-focus and track a freely moving paramecium. In this experimental result, the paramecium is auto-focused and kept inside the scope of the microscope during 45s. The evaluated focal error is within 5[μm], while a length and a thickness of the paramecium are about 200[μm] and 50[μm], respectively.

本文言語English
ホスト出版物のタイトルIROS'11 - 2011 IEEE/RSJ International Conference on Intelligent Robots and Systems
ホスト出版物のサブタイトルCelebrating 50 Years of Robotics
ページ7-12
ページ数6
DOI
出版ステータスPublished - 2011 12月 29
イベント2011 IEEE/RSJ International Conference on Intelligent Robots and Systems: Celebrating 50 Years of Robotics, IROS'11 - San Francisco, CA, United States
継続期間: 2011 9月 252011 9月 30

出版物シリーズ

名前IEEE International Conference on Intelligent Robots and Systems

Other

Other2011 IEEE/RSJ International Conference on Intelligent Robots and Systems: Celebrating 50 Years of Robotics, IROS'11
国/地域United States
CitySan Francisco, CA
Period11/9/2511/9/30

ASJC Scopus subject areas

  • 制御およびシステム工学
  • ソフトウェア
  • コンピュータ ビジョンおよびパターン認識
  • コンピュータ サイエンスの応用

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