Fully automated platelet differential interference contrast image analysis via deep learning.

Journal: Scientific reports
Published Date:

Abstract

Platelets mediate arterial thrombosis, a leading cause of myocardial infarction and stroke. During injury, platelets adhere and spread over exposed subendothelial matrix substrates of the damaged blood vessel wall. The mechanisms which govern platelet activation and their interaction with a range of substrates are therefore regularly investigated using platelet spreading assays. These assays often use differential interference contrast (DIC) microscopy to assess platelet morphology and analysis performed using manual annotation. Here, a convolutional neural network (CNN) allowed fully automated analysis of platelet spreading assays captured by DIC microscopy. The CNN was trained using 120 generalised training images. Increasing the number of training images increases the mean average precision of the CNN. The CNN performance was compared to six manual annotators. Significant variation was observed between annotators, highlighting bias when manual analysis is performed. The CNN effectively analysed platelet morphology when platelets spread over a range of substrates (CRP-XL, vWF and fibrinogen), in the presence and absence of inhibitors (dasatinib, ibrutinib and PRT-060318) and agonist (thrombin), with results consistent in quantifying spread platelet area which is comparable to published literature. The application of a CNN enables, for the first time, automated analysis of platelet spreading assays captured by DIC microscopy.

Authors

  • Carly Kempster
    School of Biological Sciences, University of Reading, Reading, UK.
  • George Butler
    School of Biological Sciences, University of Reading, Reading, UK.
  • Elina Kuznecova
    School of Biological Sciences, University of Reading, Reading, UK.
  • Kirk A Taylor
    School of Biological Sciences, University of Reading, Reading, UK.
  • Neline Kriek
    School of Biological Sciences, University of Reading, Reading, UK.
  • Gemma Little
    School of Biological Sciences, University of Reading, Reading, UK.
  • Marcin A Sowa
    School of Biological Sciences, University of Reading, Reading, UK.
  • Tanya Sage
    School of Biological Sciences, University of Reading, Reading, UK.
  • Louise J Johnson
    School of Biological Sciences, University of Reading, Reading, UK.
  • Jonathan M Gibbins
    School of Biological Sciences, University of Reading, Reading, UK.
  • Alice Y Pollitt
    School of Biological Sciences, University of Reading, Reading, UK. a.pollitt@reading.ac.uk.