Using Raman spectroscopy to assess hemoglobin oxygenation in red blood cell concentrate: an objective proxy for morphological index to gauge the quality of stored blood?

Abstract

Blood banking is an essential aspect of modern medical care. When red blood cells (RBCs) are stored, they become damaged by various chemical processes, such as accumulation of their own waste products and oxidative injury, among others. These processes lead to the development of the RBC storage lesion, a complex condition where the severity is reflected through the morphology of the stored cells. It was hypothesized that Raman spectroscopy could be used to monitor certain structural and compositional changes associated with such ageing effects and that a relationship between these features and traditional morphology (as measured using a morphology index) could be observed. The hypothesis was tested by measuring spectral features associated with hemoglobin oxygenation from dry-fixed smears and liquid RBCs for twenty-nine different donors (combined), and comparing the trends with morphological scoring from seven of these donors. After appropriately fitting the two data sets to either power or linear curves, the oxygenation state was shown to change in a manner that was donor-dependent and that closely tracked morphological changes. This study suggests Raman analysis has promise for providing a rapid and objective measure of the cell quality of stored RBCs through measurements of hemoglobin oxygenation that is comparable to traditional morphological assessment.

Diagnosing sickle cell disease and iron deficiency anemia in human blood by Raman spectroscopy

“…The difference spectra in Fig. 1 showed that both IDA and SCD presented lower intensity for the peaks at 1227, 1375, and 1406 cm −1 , which can be assigned to oxyhemoglobin [33,45,46]; the IDA presented lower intensity for the peaks at 753, 1567, 1622, and 1644 cm −1 , assigned to the hemoglobin molecule [46][47][48][49][50] and higher intensity for the peak at 1131 cm −1 , assigned to the pyrrolic ring of the porphyrin; the SCD presented lower intensity for the peak at 1004 cm −1 , assigned to aromatic ring of amino acids (phenylalanine) [51], and higher intensity for the peaks at 1215, 1358, 1526, 1547, and 1622 cm −1 , assigned to deoxyhemoglobin [45,46,50]. All of these labeled peaks showed statistically significant differences when comparing the three groups (HbA, IDA, and SCD) (one-way ANOVA test, p < 0.05).…”

“…The number of spectral features revealed by the loadings suggests specific biochemical changes caused by both types of anemia compared to HbA. The loading vector 1 (Loading 1) presents the spectral features referred to whole blood [28,41,44,46], with positive peaks at 753,1004,1215,1227,1358,1375,1406,1526,1547,1567,1622, and 1644 cm −1 assigned to blood cells (mainly erythrocytes) [28,41] and plasma constituents (mainly albumin) [44]. The Score 1 showed statistically significant differences between the groups IDA versus HbA, SCD versus HbA, and IDA versus SCD (one-way ANOVA, Tukey posttest, p < 0.05), with lower intensity for the IDA and higher intensity for the SCD.…”

“…The loading vector 2 (Loading 2) presented positive spectral features at 1004, 1375, and 1644 cm −1 , which can be assigned to hemoglobin and oxyhemoglobin [33,46,50,51], and negative spectral features at 753, 1215, 1358, 1526, and 1547 cm −1 , which can be assigned to deoxyhemoglobin [33,41,45,46,50]. The Score 2 showed statistically significant differences for the SCD versus HbA and SCD versus IDA (one-way ANOVA, Tukey posttest, p < 0.05), with positive values for the HbA and IDA and negative values for the SCD.…”

“…Changes in the shape of erythrocytes (discoid red blood cells can be transformed into echinocyte or stomatocyte shape) lead to change in the oxygen affinity, and it was demonstrated that changes in hemoglobin oxygenation lead to changes in the Raman peaks referred to hemoglobin [54]. Atkins et al [46] showed that these deoxyhemoglobin peaks increase with the time of storage of blood transfusion bags. According to this Atkins's study, the ration between the 1208 and 1222 cm −1 peaks correlated with the "morphology scoring" of the erythrocytes and can be used to monitor blood oxygenation status after long-term storage.…”

Laser Raman tweezer spectroscopy to explore the bisphenol A-induced changes in human erythrocytes

“…So the hemoglobin oxygenation state can be identied by monitoring the ratio (1209 cm À1 /1222 cm À1 ) of the deoxyHb/oxyHb bands which corresponds to methine C-H deformations. 36 The shi of hemoglobin molecules from deoxygenated state to oxygenated state in presence of BPA can be assumed from the increase in 1222 cm À1 peak intensity accompanied by the gradual decrease in 1209 cm À1 from the Fig. 3.…”