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A Petri-Dish with Micromolded Pattern as a Coordinate Indicator for Live-Cell Time Lapse Microscopy

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Abstract

Determining the position of cells in a microscope image is essential in investigating cell behaviors such as migration and differentiation. When observing cells over a time period over many ROIs (region of interests), it is difficult to reposition the sample and find the same cell once the FOV (field of view) is changed. This study describes a novel approach to obtain accurate position data in live-cell time-lapse imaging and large-area stitched images using a Petri-dish with embedded microstructures and a simple microscope. We integrated a braille-like pattern underneath the Petri-dish, a coordinate indicator for microscope image registration to simplify imaging of multiple ROIs at different time points without expensive equipment such as motorized stages or enclosed cell culture incubators. In this study, we micromolded microscopic structures with sizes between 5–10 μm spaced closely (~ 40 μm) in non-repeating pattern underneath the petri-dish. Image processing technique was used to determine the position and rotation of the petri-dish with respect to the microscope FOV. Using this methods, we demonstrated successful imaging of time-lapse images for cell differentiation and migration. Large area patterned cell cultures were imaged over several days and the images were stitched without a motorized microscope.

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Acknowledgements

This study was supported by the National Research Foundation of Korea (NRF) grant funded by the Korean government(MSIT) (No.2021R1A3B1077481, 2020R1A2C1006331) and a grant supported by the Ministry of Trade, Industry and Energy (MOTIE) and Korea Institute for Advancement of Technology (KIAT) through the International Cooperative R&D program (P0011266). This research was also supported by Hankuk University of Foreign Studies Research Fund (YC).

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Author notes

  1. Kyungwon Yun and Dohyun Park contributed equally to this work.

Authors and Affiliations

  1. Department of Mechanical Engineering, Seoul National University, Seoul, 151-744, Korea

    Kyungwon Yun, Dohyun Park, Myeongwoo Kang, Jiyoung Song & Noo Li Jeon

  2. Bio-MAX Institute, Seoul National University, Seoul, 08826, Republic of Korea

    Dohyun Park & Noo Li Jeon

  3. Division of Computer Engineering, Hankuk University of Foreign Studies, Yongin, 17035, Korea

    Yoojin Chung

  4. Envisible, Inc., Teheran-ro 13-Gil Gangnam-gu, Seoul, 135-080, Korea

    Hyunwoo Bang

  5. Institute of Advanced Machines and Design, Seoul National University, Seoul, 08826, Republic of Korea

    Noo Li Jeon

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  1. Kyungwon Yun
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  2. Dohyun Park
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  3. Myeongwoo Kang
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  4. Jiyoung Song
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  5. Yoojin Chung
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  7. Noo Li Jeon
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Correspondence to Hyunwoo Bang or Noo Li Jeon.

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Yun, K., Park, D., Kang, M. et al. A Petri-Dish with Micromolded Pattern as a Coordinate Indicator for Live-Cell Time Lapse Microscopy. BioChip J 16, 27–32 (2022). https://doi.org/10.1007/s13206-021-00039-8

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  • DOI: https://doi.org/10.1007/s13206-021-00039-8

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