A practical guide to intelligent image-activated cell sorting.

Journal: Nature protocols
PMID: 31278398

Abstract

Intelligent image-activated cell sorting (iIACS) is a machine-intelligence technology that performs real-time intelligent image-based sorting of single cells with high throughput. iIACS extends beyond the capabilities of fluorescence-activated cell sorting (FACS) from fluorescence intensity profiles of cells to multidimensional images, thereby enabling high-content sorting of cells or cell clusters with unique spatial chemical and morphological traits. Therefore, iIACS serves as an integral part of holistic single-cell analysis by enabling direct links between population-level analysis (flow cytometry), cell-level analysis (microscopy), and gene-level analysis (sequencing). Specifically, iIACS is based on a seamless integration of high-throughput cell microscopy (e.g., multicolor fluorescence imaging, bright-field imaging), cell focusing, cell sorting, and deep learning on a hybrid software-hardware data management infrastructure, enabling real-time automated operation for data acquisition, data processing, intelligent decision making, and actuation. Here, we provide a practical guide to iIACS that describes how to design, build, characterize, and use an iIACS machine. The guide includes the consideration of several important design parameters, such as throughput, sensitivity, dynamic range, image quality, sort purity, and sort yield; the development and integration of optical, microfluidic, electrical, computational, and mechanical components; and the characterization and practical usage of the integrated system. Assuming that all components are readily available, a team of several researchers experienced in optics, electronics, digital signal processing, microfluidics, mechatronics, and flow cytometry can complete this protocol in ~3 months.

Authors

  • Akihiro Isozaki
    Department of Chemistry, The University of Tokyo, Tokyo, Japan.
  • Hideharu Mikami
    Department of Chemistry, The University of Tokyo, Tokyo, Japan.
  • Kotaro Hiramatsu
    Department of Chemistry, The University of Tokyo, Tokyo, Japan.
  • Shinya Sakuma
    Department of Micro-Nano Mechanical Science and Engineering, Nagoya University, Nagoya, Aichi, Japan. arai@mech.nagoya-u.ac.jp.
  • Yusuke Kasai
    Department of Micro-Nano Mechanical Science and Engineering, Nagoya University, Nagoya, Japan.
  • Takanori Iino
    Department of Electrical Engineering and Information Systems, The University of Tokyo, Tokyo, Japan.
  • Takashi Yamano
    Laboratory of Applied Molecular Microbiology, Kyoto University, Kyoto, Japan.
  • Atsushi Yasumoto
  • Yusuke Oguchi
    Department of Biological Sciences, The University of Tokyo, Tokyo, Japan.
  • Nobutake Suzuki
    Department of Biological Sciences, The University of Tokyo, Tokyo, Japan.
  • Yoshitaka Shirasaki
    Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Bunkyo-ku, Tokyo, Japan.
  • Taichiro Endo
    ExaWizards Inc., Tokyo, Japan.
  • Takuro Ito
  • Kei Hiraki
    Department of Chemistry, The University of Tokyo, Tokyo, Japan.
  • Makoto Yamada
    aMs New Otani Clinic Osaka Japan.
  • Satoshi Matsusaka
    Clinical Research and Regional Innovation, Faculty of Medicine, University of Tsukuba, Ibaraki, Japan.
  • Takeshi Hayakawa
    Department of Precision Mechanics, Chuo University, Tokyo, Japan.
  • Hideya Fukuzawa
    Laboratory of Applied Molecular Microbiology, Kyoto University, Kyoto, Japan.
  • Yutaka Yatomi
  • Fumihito Arai
    Department of Micro-Nano Mechanical Science and Engineering, Nagoya University, Nagoya, Aichi, Japan. arai@mech.nagoya-u.ac.jp.
  • Dino Di Carlo
    2Department of Bioengineering, University of California, 420 Westwood Plaza, 5121 Engineering V, PO Box 951600, Los Angeles, CA 90095 USA.
  • Atsuhiro Nakagawa
  • Yu Hoshino
    Department of Chemical Engineering, Kyushu University, Fukuoka, Japan.
  • Yoichiroh Hosokawa
    Division of Materials Science, Graduate School of Science and Technology, Nara Institute of Science and Technology, Ikoma, Japan.
  • Sotaro Uemura
    Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Bunkyo-ku, Tokyo, Japan.
  • Takeaki Sugimura
    Department of Chemistry, The University of Tokyo, Tokyo, Japan.
  • Yasuyuki Ozeki
  • Nao Nitta
  • Keisuke Goda

Keywords

External Resources

Popular Topics
Recent Journals