JRM Vol.25 No.1 pp. 115-124
doi: 10.20965/jrm.2013.p0115


High-Speed Focusing and Tracking of Multisized Microbiological Objects

Chanh-Nghiem Nguyen, Kenichi Ohara, Yasushi Mae,
and Tatsuo Arai

Department of Systems Innovation, Graduate School of Engineering Science, Osaka University, 1-3 Machikaneyama, Toyonaka, Osaka 560-8531, Japan

May 23, 2012
June 5, 2012
February 20, 2013
high-speed tracking, multisized, microobject

This paper proposes a novel algorithm for high-speed autofocusing and tracking of multisized microbiological objects observed under a transmitted light microscope. Unlike well-known autofocus algorithms found in the literature, the intensity variation of only a small defined region around the border of the microobject is analyzed in the frequency domain to determine the focused position of the object quickly. In the experiment, 20 µm3T3-SWISS cells were used as smallermicroobjects and 97 µm diameter microspheres were used to represent larger microbiological objects. The execution time and accuracy of the proposed algorithmwere assessed and better results were obtained compared to some related autofocusing algorithms. Since its computational cost was low, the algorithm facilitated highspeed autofocusing of both 3T3-SWISS cells and microspheres. The algorithm was also applied to the tracking of moving microobjects by implementing a PD controller. Since visual feedback took only about 1 ms, high-speed tracking was achieved.

Cite this article as:
C. Nguyen, K. Ohara, Y. Mae, and <. Arai, “High-Speed Focusing and Tracking of Multisized Microbiological Objects,” J. Robot. Mechatron., Vol.25, No.1, pp. 115-124, 2013.
Data files:
  1. [1] F. C. Groen, I. T. Young, and G. Ligthart, “A Comparison of Different Focus Functions for Use in Autofocus Algorithms,” Cytometry, Vol.6, No.2, pp. 81-91, 1985.
  2. [2] Y. Sun, S. Duthaler, and B. J. Nelson, “Autofocusing in Computer microscopy: Selecting the Optimal Focus Algorithm,” Microscopy Research and Technique, Vol.65, No.3, pp. 139-149, 2004.
  3. [3] J. M. Mateos-Pérez, R. Redondo, R. Nava, J. C. Valdiviezo, G. Cristóbal, B. Escalante-Ramírez, M. J. Ruiz-Serrano, J. Pascau, and M. Desco, “Comparative Evaluation of Autofocus Algorithms for a Real-Time System for Automatic Detection of Mycobacterium Tuberculosis,” Cytometry Part A, Vol.81A, No.3, pp. 213-221, 2012.
  4. [4] V. V. Bezzubik and S. N. Ustinov, “Optimization of Algorithms for Autofocusing a Digital Microscope,” J. Opt. Technol., Vol.76, No.10, pp. 603-608, 2009.
  5. [5] M. A. Bueno-Ibarra, J. Álvarez-Borrego, L. Acho, and M. C. Chávez-Sánchez, “Fast Autofocus Algorithm for Automated Microscopes,” Optical Engineering, Vol.44, No.6, p. 063601, 2005.
  6. [6] K. Ohba, C. Ortega, K. Tanie, G. Rin, R. Dangi, Y. Takei, T. Kaneko, and N. Kawahara, “Real-Time Micro Observation Technique for Tele-Micro-Operation,” in IEEE/RSJ Int. Conf. on Intelligent Robots and Systems, Vol.1, pp. 647-652, 2000.
  7. [7] K. Ohara, K. Ohba, T. Tanikawa, M. Hiraki, S. Wakatsuki, and M. Mizukawa, “Hands Free Micro Operation for Protein Crystal Analysis,” in IEEE/RSJ Int. Conf. on Intelligent Robots and Systems, Vol.2, pp. 1728-1733, 2004.
  8. [8] C.-N. Nguyen, K. Ohara, E. Avci, T. Takubo, Y. Mae, and T. Arai, “Automated Micromanipulation of a Microhand with All-In-Focus Imaging System,” IEEE/RSJ Int. Conf. on Intelligent Robots and Systems, pp. 427-432, Sept. 25-30, 2011.
  9. [9] Y. Nakabo, M. Ishikawa, H. Toyoda, and S. Mizuno, “1 ms Column Parallel Vision System and Its Application of High Speed Target Tracking,” in IEEE Int. Conf. on Robotics and Automation, Vol.1, pp. 650-655, 2000.
  10. [10] H. Oku and M. Ishikawa, “High-Speed Autofocusing of a Cell Using Diffraction Pattern,” Optics Express, Vol.14, No.9, pp. 3952-3960, 2006.
  11. [11] T. Hasegawa, N. Ogawa, H. Oku, and M. Ishikawa, “A New Framework for Microrobotic Control of Motile Cells Based on High-Speed Tracking and Focusing,” IEEE Int. Conf. on Robotics and Automation, pp. 3964-3969, 2008.
  12. [12] T. Obara, Y. Igarashi, and K. Hashimoto, “Fast and Adaptive Auto-Focusing Algorithm for Microscopic Cell Observation,” in IEEE/RSJ Int. Conf. on Intelligent Robots and Systems, pp. 7-12, Sept. 25-30, 2011.
  13. [13] H. Sasaki, K. Nomura, H. Nakajima, and K. Kobayashi, “Tracking of Moving Object by Phase-only Correlation,” J. of Robotics and Mechatronics, Vol.12, No.5, pp. 541-544, 2000.
  14. [14] M. Hashimoto, Y. Matsui, and K. Takahashi, “Moving-Object Tracking with In-Vehicle Multi-Laser Range Sensors,” J. of Robotics and Mechatronics, Vol.20, No.3, pp. 367-377, 2008.
  15. [15] K. Shimonomura, K. Inoue, S. Kameda, and T. Yagi, “A Novel Robot Vision Applicable to Real-time Target Tracking,” J. of Robotics and Mechatronics, Vol.15, No.2, pp. 185-191, 2003.
  16. [16] N. Ogawa, H. Oku, K. Hashimoto, and M. Ishikawa, “Trajectory Planning of Motile Cell for Microrobotic Applications,” J. of Robotics and Mechatronics, Vol.19, No.2, pp. 190-197, 2007.
  17. [17] Y. Watanabe, T. Komuro, S. Kagami, and M. Ishikawa, “Multitarget tracking using a vision chip and its applications to real-time visual measurements,” J. of Robotics and Mechatronics, Vol.17, No.2, pp. 121-129, 2005.
  18. [18] J. M. Geusebroek, F. Cornelissen, A. W. Smeulders, and H. Geerts, “Robust Autofocusing in Microscopy,” Cytometry, Vol.39, pp. 1-9, 2000.
  19. [19] M. Frigo, and S. G. Johnson, “The Design and Implementation of FFTW3,” Proc. of the IEEE, Vol.93, No.2, pp. 216-231, 2005.

*This site is desgined based on HTML5 and CSS3 for modern browsers, e.g. Chrome, Firefox, Safari, Edge, IE9,10,11, Opera.

Last updated on Apr. 22, 2019