Abstract
Distortions of the normal bi-concave disc shape for red blood cells (RBCs) appear in a number of pathologies resulting from defects in cell membrane skeletal architecture, erythrocyte ageing, and mechanical damage. We present here the potential of acoustic cytometry for developing new approaches to light-scattering based evaluation of red blood cell disorders and of the effects of storage and ageing on changes or damage to RBCs membranes. These approaches could be used to immediately evaluate the quality of erythrocytes prior to blood donation and following transfusion. They could also be applied to studying RBC health in diseases and other pathologies, such as artificial heart valve hemolysis, thermal damage or osmotic fragility. Abnormal distributions of erythrocytes can typically be detected after just 30 to 45 seconds of acquisition time using 1–2 µL starting blood volumes.
Highlights
In 1977, Mack Fulwyler[2] demonstrated hydrodynamic cell orientation with chicken erythrocytes and he suggested that controlling the orientation of non-spherical particles would improve precision of analysis in flow cytometry
After noticing unusual arch-shaped patterns for red blood cells (RBCs) on forward vs. side scatter plots of diluted whole blood collected on the acoustic focusing cytometer, experiments were run to determine how unique these patterns are to this instrument and whether the acoustic focusing in the sample injector was playing a role
Focused instrument A showed a single diffuse RBC population, having an overall side scatter distribution covering an order of magnitude in intensity and a forward scatter distribution approximately 3 times smaller
Summary
We present here the potential of acoustic cytometry for developing new approaches to light-scattering based evaluation of red blood cell disorders and of the effects of storage and ageing on changes or damage to RBCs membranes. These approaches could be used to immediately evaluate the quality of erythrocytes prior to blood donation and following transfusion. The precision of flow cytometry measurements for non-spherical cells such as sperm cells and red blood cells is affected by the orientation of cells as they pass through a light source This is especially true for light scatter measurements, where nearly identically shaped cells can have significantly different scatter signals owing to the variation in the cross section of each individual cell facing a focused laser source. These differences are indicative of each of the conditions tested, and they show promise that this method could be developed for reagent free study of red blood cell (RBC) health and pathology
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