CELL COUNTS USING CELLPROFILER FREE

Traditional flow cytometers classify stained cells in a high-throughput, low-content manner based on a small number of light-scatter properties (forward scatter and side scatter), and fluorescence intensity features. This procedure is widely adopted in the clinical routine with well-established standard laboratory protocols. The reference range for WBC differential count is as follows: lymphocytes (20вЂ“40%), eosinophils (1вЂ“6%), monocytes (2вЂ“10%), and neutrophils (40вЂ“80%) ( ).Д¬onventionally, WBC differential count often involves flow cytometry using fluorescently tagged antibodies that are known to differentially label the WBC populations. These WBC populations have characteristic concentration ranges in healthy persons deviations, whether high or low, are clinically significant 1. They are categorized into five main types: lymphocytes (including B and T cells), eosinophils, neutrophils, monocytes, and basophils. WBCs play an important role in the body's immune system and in the defense against infections. White blood cell (WBC) differential count is a clinical test that measures the number and percentage of each WBC type in a person's sample of blood. on behalf of International Society for Advancement of Cytometry. Cytometry Part A published by Wiley Periodicals, Inc. It also opens the door to employing machine learning for liquid biopsy, here, using the rich information in cell morphology for a wide range of diagnostics of primary blood.

CELL COUNTS USING CELLPROFILER FREE

The presented method enables robust and highly accurate identification of WBCs, minimizing the disturbance to the cells and leaving marker channels free to answer other biological questions. We validated the WBC analysis with unstained samples from 85 donors. We provide an open-source workflow to carry out the procedure. It achieved an average F1-score of 97% and two subtypes of WBCs, B and T lymphocytes, were distinguished from each other with an average F1-score of 78%, a task previously considered impossible for unlabeled samples. We present a novel label-free approach using an imaging flow cytometer and machine learning algorithms, where live, unstained WBCs were classified. However, this process requires several sample preparation steps and may adversely disturb the cells. Fluorescent markers and a flow cytometer are required for the current state-of-the-art method for determining WBC differential counts. White blood cell (WBC) differential counting is an established clinical routine to assess patient immune system status.

Cell counts using cellprofiler