Classification of Five Cell Types from PBMC Samples using Single Cell Transcriptomics and Artificial Neural Networks We used 27 human single cell transcriptomics (SCT) data sets to develop an artificial neural network (ANN) model for classification of Peripheral Blood Mononuclear Cells (PBMC). We demonstrated that highly accurate models for the classification of PBMC subtypes can be developed by combining multiple independent data sets to form training data sets. A significant data preparation effort was needed for building predictive models. Using a data set of xe2x88xbc120,000 single cell instances we showed the accuracy of classification of PBMC call of xe2x88xbc 90%. Optimization techniques and the addition of new highquality data sets for model training are expected to improve PBMC subtype classification accuracy.

Classification of human white blood cells using machine learning for stainfree imaging flow cytometry Imaging flow cytometry (IFC) produces up to 12 different informationrich images of single cells at a throughput of 5000 cells per second. Yet often, cell populations are still studied using manual gating, a technique that has several drawbacks. Firstly, it is hard to reproduce. Secondly, it is subjective and biased. And thirdly, it is timeconsuming for large experiments. Therefore, it would be advantageous to replace manual gating with an automated process, which could be based on stainfree measurements originating from the brightfield and darkfield image channels. To realise this potential, advanced data analysis methods are required, in particular, machine learning. Previous works have successfully tested this approach on cell cycle phase classification with both a classical machine learning approach based on manually engineered features, and a deep learning approach. In this work, we compare both approaches extensively on the complex problem of white blood cell classification. Four human whole blood samples were assayed on an ImageStreamX MK II imaging flow cytometer. Two samples were stained for the identification of 8 white blood cell types, while two other sample sets were stained for the identification of resting and active eosinophils. For both datasets, four machine learning classifiers were evaluated on stainfree imagery using stratified 5fold crossvalidation. On the white blood cell dataset the best obtained results were 0.776 and 0.697 balanced accuracy for classical machine learning and deep learning, respectively. On the eosinophil dataset this was 0.866 and 0.867 balanced accuracy. From the experiments we conclude that classifying distinct cell types based on only stainfree images is possible with these techniques. However, both approaches did not always succeed in making reliable cell subtype classifications. Also, depending on the cell type, we find that even though the deep learning approach requires less expert input, it performs on par with a classical approach.