Premium
Quantifying morphological heterogeneity: a study of more than 1 000 000 individual stored red blood cells
Author(s) -
Piety N. Z.,
Gifford S. C.,
Yang X.,
Shevkoplyas S. S.
Publication year - 2015
Publication title -
vox sanguinis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.68
H-Index - 83
eISSN - 1423-0410
pISSN - 0042-9007
DOI - 10.1111/vox.12277
Subject(s) - blood preservation , biology , evolutionary biology , physiology
Background and Objectives The morphology of red blood cells ( RBC s) 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 RBC s are manual, laborious and 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 RBC s, 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 RBC s 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 RBC s (speed: >10 000 cells/h; accuracy: 91·9% low resolution, 75·3% high resolution). Diameter mean and standard deviation by morphology class were as follows: discocyte 7·80 ± 0·49 μm, echinocyte 1 7·61 ± 0·63 μm, echinocyte 2 7·02 ± 0·61 μm, echinocyte 3 6·47 ± 0·42 μm, sphero‐echinocyte 6·01 ± 0·26 μm, spherocyte 6·02 ± 0·27 μm, stomatocyte 1 6·95 ± 0·61 μm and stomatocyte 2 7·32 ± 0·47 μm. Conclusions 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 RBC s automatically, rapidly and in statistically significant numbers enabled us to perform the most extensive study of stored RBC morphology to date.