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Raman spectroscopy of transfusable red blood cells Atkins, Chad Garry

Abstract

Blood banking is an essential aspect of modern healthcare. When red blood cells (RBCs) are stored, they degrade over time as a result of various chemical and biological processes leading to an accumulation of waste products and oxidative damage, among others. Significant growth in the application of Raman spectroscopy (RS) to biomedical problems has made it a feasible tool for investigating biochemical changes associated with storage of RBCs. It was hypothesized that RS could be used to monitor in situ, non-destructively and non-invasively, certain structural and compositional changes associated with these ageing effects as they occur in storage bags. The presence of a relationship between these changes and the viability of RBCs would have substantial implications for the health care industry. Preliminary results demonstrated the oxygenation state of hemoglobin in stored RBCs changed in a manner that was donor-dependent and that closely tracked the morphological index, a qualitative metric for evaluating cell quality. Investigations of the storage-solution supernatant revealed that lactate, a metabolic waste product, accumulated at different rates in stored bags and displayed more rapid accumulation in units from male donors than units from female donors. It was shown that spatially offset Raman spectroscopy (SORS) could measure stored RBC biochemistry through the plastic bag. Spectra collected with SORS compared well with previously-obtained conventional Raman spectra. Thus, information about the contents of a stored unit could be obtained without breaching its sterility. These outcomes provided proof-of-concept for the development of a SORS-based instrument to measure storage bag contents in a rapid and non-invasive manner. The results reported in this thesis fall into two distinct categories. Regarding the observed data, the substantial inter-donor variability and gender and age effects have significant implications for the design of clinical trials, the collection and administration of donated blood, and clinical use. Regarding instrumentation, obtaining chemical information from stored RBCs using RS will enable new research directions for the field of transfusion medicine and obtaining this information in situ from storage bags will have utility for quality control in a blood bank or hospital setting.

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Attribution-NonCommercial-NoDerivatives 4.0 International