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Automated spheroid generation, drug application and efficacy screening using a deep learning classification: a feasibility study.

ABSTRACT: The last two decades saw the establishment of three-dimensional (3D) cell cultures as an acknowledged tool to investigate cell behaviour in a tissue-like environment. Cells growing in spheroids differentiate and develop different characteristics in comparison to their two-dimensionally grown counterparts and are hence seen to exhibit a more in vivo-like phenotype. However, generating, treating and analysing spheroids in high quantities remains labour intensive and therefore limits its applicability in drugs and compound research. Here we present a fully automated pipetting robot that is able to (a) seed hanging drops from single cell suspensions, (b) treat the spheroids formed in these hanging drops with drugs and (c) analyse the viability of the spheroids by an image-based deep learning based convolutional neuronal network (CNN). The model is trained to classify between ‘unaffected’, ‘mildly affected’ and ‘affected’ spheroids after drug exposure. All corresponding spheroids are initially analysed by viability flow cytometry analysis to build a labelled training set for the CNN to subsequently reduce the number of misclassifications. Hence, this approach allows to efficiently examine the efficacy of drug combinatorics or new compounds in 3D cell culture. Additionally, it may provide a valuable instrument to screen for new and individualized systemic therapeutic strategies in second and third line treatment of solid malignancies using patient derived primary cells.

SUBMITTER: Benning L

PROVIDER: S-EPMC7338379 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature

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Project description:The COVID-19 pandemic has caused a worldwide catastrophe and widespread devastation that reeled almost all countries. The pandemic has mounted pressure on the existing healthcare system and caused panic and desperation. The gold testing standard for COVID-19 detection, reverse transcription-polymerase chain reaction (RT-PCR), has shown its limitations with 70% accuracy, contributing to the incorrect diagnosis that exaggerated the complexities and increased the fatalities. The new variations further pose unseen challenges in terms of their diagnosis and subsequent treatment. The COVID-19 virus heavily impacts the lungs and fills the air sacs with fluid causing pneumonia. Thus, chest X-ray inspection is a viable option if the inspection detects COVID-19-induced pneumonia, hence confirming the exposure of COVID-19. Artificial intelligence and machine learning techniques are capable of examining chest X-rays in order to detect patterns that can confirm the presence of COVID-19-induced pneumonia. This research used CNN and deep learning techniques to detect COVID-19-induced pneumonia from chest X-rays. Transfer learning with fine-tuning ensures that the proposed work successfully classifies COVID-19-induced pneumonia, regular pneumonia, and normal conditions. Xception, Visual Geometry Group 16, and Visual Geometry Group 19 are used to realize transfer learning. The experimental results were promising in terms of precision, recall, F1 score, specificity, false omission rate, false negative rate, false positive rate, and false discovery rate with a COVID-19-induced pneumonia detection accuracy of 98%. Experimental results also revealed that the proposed work has not only correctly identified COVID-19 exposure but also made a distinction between COVID-19-induced pneumonia and regular pneumonia, as the latter is a very common disease, while COVID-19 is more lethal. These results mitigated the concern and overlap in the diagnosis of COVID-19-induced pneumonia and regular pneumonia. With further integrations, it can be employed as a potential standard model in differentiating the various lung-related infections, including COVID-19.

Project description:PurposeTo develop deep learning models to perform automated diagnosis and quantitative classification of age-related cataract from anterior segment photographs.DesignDeepLensNet was trained by applying deep learning models to the Age-Related Eye Disease Study (AREDS) dataset.ParticipantsA total of 18 999 photographs (6333 triplets) from longitudinal follow-up of 1137 eyes (576 AREDS participants).MethodsDeep learning models were trained to detect and quantify nuclear sclerosis (NS; scale 0.9-7.1) from 45-degree slit-lamp photographs and cortical lens opacity (CLO; scale 0%-100%) and posterior subcapsular cataract (PSC; scale 0%-100%) from retroillumination photographs. DeepLensNet performance was compared with that of 14 ophthalmologists and 24 medical students.Main outcome measuresMean squared error (MSE).ResultsOn the full test set, mean MSE for DeepLensNet was 0.23 (standard deviation [SD], 0.01) for NS, 13.1 (SD, 1.6) for CLO, and 16.6 (SD, 2.4) for PSC. On a subset of the test set (substantially enriched for positive cases of CLO and PSC), for NS, mean MSE for DeepLensNet was 0.23 (SD, 0.02), compared with 0.98 (SD, 0.24; P = 0.000001) for the ophthalmologists and 1.24 (SD, 0.34; P = 0.000005) for the medical students. For CLO, mean MSE was 53.5 (SD, 14.8), compared with 134.9 (SD, 89.9; P = 0.003) for the ophthalmologists and 433.6 (SD, 962.1; P = 0.0007) for the medical students. For PSC, mean MSE was 171.9 (SD, 38.9), compared with 176.8 (SD, 98.0; P = 0.67) for the ophthalmologists and 398.2 (SD, 645.4; P = 0.18) for the medical students. In external validation on the Singapore Malay Eye Study (sampled to reflect the cataract severity distribution in AREDS), the MSE for DeepSeeNet was 1.27 for NS and 25.5 for PSC.ConclusionsDeepLensNet performed automated and quantitative classification of cataract severity for all 3 types of age-related cataract. For the 2 most common types (NS and CLO), the accuracy was significantly superior to that of ophthalmologists; for the least common type (PSC), it was similar. DeepLensNet may have wide potential applications in both clinical and research domains. In the future, such approaches may increase the accessibility of cataract assessment globally. The code and models are available at https://github.com/ncbi/deeplensnet.

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Automated spheroid generation, drug application and efficacy screening using a deep learning classification: a feasibility study.

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