Interpreting lymphocyte classification from Hoechst stained slides with deep learning

Jessica Cooper*, In Hwa Um, Oggie Arandelovic, David James Harrison

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

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Multiplex immunofluorescence and immunohistochemistry benefit patients by allowing cancer pathologists to identify proteins expressed on the surface of cells. This enables cell classification, better understanding of the tumour microenvironment, and more accurate diagnoses, prognoses, and tailored immunotherapy based on the immune status of individual patients. However, these techniques are expensive. They are time consuming processes which require complex staining and imaging techniques by expert technicians. Hoechst staining is far cheaper and easier to perform, but is not typically used as it binds to DNA rather than to the proteins targeted by immunofluorescence techniques. In this work we show that through the use of deep learning it is possible to identify an immune cell subtype without immunofluorescence. We train a deep convolutional neural network to identify cells expressing the T lymphocyte marker CD3 from Hoechst 33342 stained tissue only. CD3 expressing cells are often used in key prognostic metrics such as assessment of immune cell infiltration, and by identifying them without the need for costly immunofluorescence, we present a promising new approach to cheaper prediction and improvement of patient outcomes. We also show that by using deep learning interpretability techniques, we can gain insight into the previously unknown morphological features which make this possible.
Original languageEnglish
Article number5957
Number of pages9
Issue number23
Early online date2 Dec 2022
Publication statusPublished - 2 Dec 2022


  • Deep learning
  • Computer vision
  • Lymphocyte subsets
  • Image classification
  • Imaging


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