Quantifying morphological heterogeneity: a study of more than 1 000 000 individual stored red blood cells


Background and Objectives The morphology of red blood cells (RBCs) deteriorates progressively during hypothermic storage. The degree of deterioration varies between individual cells, resulting in a highly heterogeneous population of cells contained within each RBC unit. Current techniques capable of categorizing the morphology of individual stored RBCs are manual, laborious, error-prone procedures that limit the number of cells that can be studied. Our objective was to create a simple, automated system for high-throughput RBC morphology classification.

Materials and Methods A simple microfluidic device, designed to enable rapid, consistent acquisition of images of optimally oriented RBCs, was fabricated using soft lithography. A custom image analysis algorithm was developed to categorize the morphology of each individual RBC in the acquired images. The system was used to determine morphology of individual RBCs in several RBC units stored hypothermically for 6–8 weeks.

Results The system was used to automatically determine the distribution of cell diameter within each morphological class for >1,000,000 individual stored RBCs (speed: >10,000 cells/hour; accuracy: 91.9% low-resolution, 75.3% high-resolution). Diameter mean and standard deviation by morphology class: discocyte 7.80±0.49?m, echinocyte 1 7.61±0.63?m, echinocyte 2 7.02±0.61?m, echinocyte 3 6.47±0.42?m, sphero-echinocyte 6.01±0.26?m, spherocyte 6.02±0.27?m, stomatocyte 1 6.95±0.61?m, stomatocyte 2 7.32 ± 0.47?m.

Conclusion The automated morphology classification procedure described in this study is significantly simpler, faster and less subjective than conventional manual procedures. The ability to evaluate the morphology of individual RBCs automatically, rapidly and in statistically significant numbers enabled us to perform the most extensive study of stored RBC morphology to date.



storage, morphology, microfluidic


Copyright 2015 Vox Sanguinis. This is a post-print version of a published paper that is available at: https://onlinelibrary.wiley.com/doi/full/10.1111/vox.12277 Recommended citation: Piety, N. Z., S. C. Gifford, X. Yang, and S. S. Shevkoplyas. "Quantifying morphological heterogeneity: a study of more than 1 000 000 individual stored red blood cells." Vox sanguinis 109, no. 3 (2015): 221-230. DOI: 10.1111/vox.12277 This item has been deposited in accordance with publisher copyright and licensing terms and with the author’s permission.