Cheetah: A Computational Toolkit for Cybergenetic Control.

Journal: ACS synthetic biology
PMID:

Abstract

Advances in microscopy, microfluidics, and optogenetics enable single-cell monitoring and environmental regulation and offer the means to control cellular phenotypes. The development of such systems is challenging and often results in bespoke setups that hinder reproducibility. To address this, we introduce Cheetah, a flexible computational toolkit that simplifies the integration of real-time microscopy analysis with algorithms for cellular control. Central to the platform is an image segmentation system based on the versatile U-Net convolutional neural network. This is supplemented with functionality to robustly count, characterize, and control cells over time. We demonstrate Cheetah's core capabilities by analyzing long-term bacterial and mammalian cell growth and by dynamically controlling protein expression in mammalian cells. In all cases, Cheetah's segmentation accuracy exceeds that of a commonly used thresholding-based method, allowing for more accurate control signals to be generated. Availability of this easy-to-use platform will make control engineering techniques more accessible and offer new ways to probe and manipulate living cells.

Authors

  • Elisa Pedone
    Department of Engineering Mathematics, University of Bristol, Ada Lovelace Building, University Walk, BS8 1TW Bristol, United Kingdom.
  • Irene de Cesare
    Department of Engineering Mathematics, University of Bristol, Ada Lovelace Building, University Walk, BS8 1TW Bristol, United Kingdom.
  • Criseida G Zamora-Chimal
    Department of Engineering Mathematics, University of Bristol, Ada Lovelace Building, University Walk, BS8 1TW Bristol, United Kingdom.
  • David Haener
    Department of Engineering Mathematics, University of Bristol, Ada Lovelace Building, University Walk, BS8 1TW Bristol, United Kingdom.
  • Lorena Postiglione
    Department of Engineering Mathematics, University of Bristol, Ada Lovelace Building, University Walk, BS8 1TW Bristol, United Kingdom.
  • Antonella La Regina
    Department of Engineering Mathematics, University of Bristol, Ada Lovelace Building, University Walk, BS8 1TW Bristol, United Kingdom.
  • Barbara Shannon
    BrisSynBio, Life Sciences Building, Tyndall Avenue, BS8 1TQ Bristol, United Kingdom.
  • Nigel J Savery
    BrisSynBio, Life Sciences Building, Tyndall Avenue, BS8 1TQ Bristol, United Kingdom.
  • Claire S Grierson
    BrisSynBio, Life Sciences Building, Tyndall Avenue, BS8 1TQ Bristol, United Kingdom.
  • Mario Di Bernardo
    Department of Engineering Mathematics, University of Bristol, Bristol, United Kingdom.
  • Thomas E Gorochowski
    School of Biological Sciences, University of Bristol, Life Sciences Building Tyndall Avenue, Bristol BS8 1TQ, UK; BrisSynBio, University of Bristol, Life Sciences Building, Tyndall Avenue, Bristol BS8 1TQ, UK. Electronic address: thomas.gorochowski@bristol.ac.uk.
  • Lucia Marucci
    Department of Engineering Mathematics, University of Bristol, Ada Lovelace Building, University Walk, BS8 1TW Bristol, United Kingdom.