Multiplexed high-throughput localized electroporation workflow with deep learning-based analysis for cell engineering.

Journal: Science advances
PMID: 35867793

Abstract

Manipulation of cells for applications such as biomanufacturing and cell-based therapeutics involves introducing biomolecular cargoes into cells. However, successful delivery is a function of multiple experimental factors requiring several rounds of optimization. Here, we present a high-throughput multiwell-format localized electroporation device (LEPD) assisted by deep learning image analysis that enables quick optimization of experimental factors for efficient delivery. We showcase the versatility of the LEPD platform by successfully delivering biomolecules into different types of adherent and suspension cells. We also demonstrate multicargo delivery with tight dosage distribution and precise ratiometric control. Furthermore, we used the platform to achieve functional gene knockdown in human induced pluripotent stem cells and used the deep learning framework to analyze protein expression along with changes in cell morphology. Overall, we present a workflow that enables combinatorial experiments and rapid analysis for the optimization of intracellular delivery protocols required for genetic manipulation.

Authors

  • Cesar A Patino
    Department of Mechanical Engineering, Northwestern University, Evanston, IL, USA.
  • Nibir Pathak
    Department of Mechanical Engineering, Northwestern University, Evanston, IL, USA.
  • Prithvijit Mukherjee
    Department of Mechanical Engineering, Northwestern University, Evanston, IL, USA.
  • So Hyun Park
    Department of Bioengineering, Rice University, 6500 Main St, Houston, TX 77030, USA.
  • Gang Bao
    Hubei Key Laboratory of Cascaded Hydropower Stations Operation & Control, Electrical Engineering & New Energy, China Three Gorges University, Yichang 443002, China. Electronic address: ctgugangbao@ctgu.edu.cn.
  • Horacio D Espinosa
    Department of Mechanical Engineering, Northwestern University, Evanston, IL, USA.

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