Raman characterizations of red blood cells with β-thalassemia using laser tweezers Raman spectroscopy

This study aimed to study the differences in Raman spectra of red blood cells (RBCs) among patients with β-thalassemia and controls using laser tweezers Raman spectroscopy (LTRS) system. A total of 33 patients with β-thalassemia major, 49 with β-thalassemia minor, and 65 controls were studied. Raman spectra of RBCs for each sample were recorded. Principal component analysis (PCA), one-way analysis of variance (ANOVA), and independent-sample t test were performed. The intensities of Raman spectra of β-thalassemia (major and minor) RBCs were lower than those of controls, especially at bands 1546, 1603, and 1619 cm(–1). The intensity ratio of band 1546 cm(–1) to band 1448 cm(–1) demonstrated that there was a significant difference between the spectra of β-thalassemia major (mostly below 2.15) and those of controls. The spectra of controls could be well distinguished from those of β-thalassemia major using PCA. After normalization, the spectra of two different genotypes with β(0)/β(0) mutations mainly overlapped, while those with β(+)/β(+) mutations had lower intensity at bands 1546, 1603, and 1619 cm(–1). The present study provided Raman characteristics of RBCs in patients with β-thalassemia major and supported the use of LTRS as a method for screening β-thalassemia major. The recognition rate for β-thalassemia minor needs to be further improved.