Prediction of Spheroid Cell Death Using Fluorescence Staining and Convolutional Neural Networks

Srisongkram, Tarapong; Syahid, Nur Fadhilah; Piyasawetkul, Thanawat; Thirawatthanasak, Pannaphat; Khamtang, Patcharapa; Sawasnopparat, Nathida; Tookkane, Dheerapat; Weerapreeyakul, Natthida; Puthongking, Ploenthip

doi:10.1021/acs.chemrestox.3c00257

Chem. Res. Toxicol.

2023

DOI: 10.1021/acs.chemrestox.3c00257

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Prediction of Spheroid Cell Death Using Fluorescence Staining and Convolutional Neural Networks

Tarapong Srisongkram,

Nur Fadhilah Syahid,

Thanawat Piyasawetkul

et al.

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2024

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Cited by 1 publication

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High-throughput platform for label-free sorting of 3D spheroids using deep learning

Sampaio da Silva,

Boos,

Goldowsky

et al. 2024

Front. Bioeng. Biotechnol.

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End-stage liver diseases have an increasing impact worldwide, exacerbated by the shortage of transplantable organs. Recognized as one of the promising solutions, tissue engineering aims at recreating functional tissues and organs in vitro. The integration of bioprinting technologies with biological 3D models, such as multi-cellular spheroids, has enabled the fabrication of tissue constructs that better mimic complex structures and in vivo functionality of organs. However, the lack of methods for large-scale production of homogeneous spheroids has hindered the upscaling of tissue fabrication. In this work, we introduce a fully automated platform, designed for high-throughput sorting of 3D spheroids based on label-free analysis of brightfield images. The compact platform is compatible with standard biosafety cabinets and includes a custom-made microscope and two fluidic systems that optimize single spheroid handling to enhance sorting speed. We use machine learning to classify spheroids based on their bioprinting compatibility. This approach enables complex morphological analysis, including assessing spheroid viability, without relying on invasive fluorescent labels. Furthermore, we demonstrate the efficacy of transfer learning for biological applications, for which acquiring large datasets remains challenging. Utilizing this platform, we efficiently sort mono-cellular and multi-cellular liver spheroids, the latter being used in bioprinting applications, and confirm that the sorting process preserves viability and functionality of the spheroids. By ensuring spheroid homogeneity, our sorting platform paves the way for standardized and scalable tissue fabrication, advancing regenerative medicine applications.

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High-throughput platform for label-free sorting of 3D spheroids using deep learning

Sampaio da Silva,

Boos,

Goldowsky

et al. 2024

Front. Bioeng. Biotechnol.

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show abstract

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Prediction of Spheroid Cell Death Using Fluorescence Staining and Convolutional Neural Networks

Cited by 1 publication

References 27 publications

High-throughput platform for label-free sorting of 3D spheroids using deep learning

High-throughput platform for label-free sorting of 3D spheroids using deep learning

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