Unraveling biochemical differences in the membrane of functional RBCs and elliptocytes using vortex beam-based micro-Raman spectroscopy.

Understanding the complexity of membrane biochemical changes in in-vitro-induced elliptocytosis can be interesting as it may mimic those in hereditary elliptocytosis. Studying the membrane biochemical changes in metabolically active elliptocytes can be crucial, but most modern methods, such as ektacytometry and EMA binding tests, fail to do so. This study employs single-cell Raman spectroscopy, a proven technique to study biochemical changes in individual functional cells to investigate biochemical modifications in the membrane and cytoskeleton of elliptocytes. This was possible by applying a vortex beam, which can probe the RBC membrane with a reduced contribution from hemoglobin, which otherwise dominates the cell spectrum. Raman spectral variations in elliptocytes indicated changes in proteins, lipids, and lipid-protein interactions. The study also presented an incidental observation of diversity in membrane components and membrane-hemoglobin interaction among tested individuals.