Browsing by Author "Lush, Richard"
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Item Open Access FT-infrared spectroscopic studies of lymphoma, lymphoid and myeloid leukaemia cell lines(2007-12-31T00:00:00Z) Babrah, Jaspreet; McCarthy, Keith P.; Lush, Richard; Rye, Adam D.; Bessant, Conrad M.; Stone, Nicholas; Dietrich, Schweitzer; Maryann, FitzmauriceThis paper presents a novel method to characterise spectral differences that distinguish leukaemia and lymphoma cell lines. This is based on objective spectral measurements of major cellular biochemical constituents and multivariate spectral processing. Fourier transform infrared (FT-IR) maps of the lymphoma, lymphoid and myeloid leukaemia cell samples were obtained using a Perkin-Elmer Spotlight 300 FT-IR imaging spectrometer. Multivariate statistical techniques incorporating principal component analysis (PCA) and linear discriminant analysis (LDA) were used to construct a mathematical model. This model was validated for reproducibility. Multivariate statistical analysis of FTIR spectra collected for each cell sample permit a combination of unsupervised and supervised methods of distinguishing cell line types. This resulted in the clustering of cell line populations, indicating distinct bio-molecular differences. Major spectral differences were observed in the 4000 to 800 cm- 1 spectral region. Bands in the averaged spectra for the cell line were assigned to the major biochemical constituents including; proteins, fatty acids, carbohydrates and nucleic acids. The combination of FT-IR spectroscopy and multivariate statistical analysis provides an important insight into the fundamental spectral differences between the cell lines, which differ according to the cellular biochemical composition. These spectral differences can serve as potential biomarkers for the differentiation of leukaemia and lymphoma cells. Consequently these differences could be used as the basis for developing a spectral method for the detection and identification of haematological malignancies.Item Open Access Vibrational spectroscopy for the rapid and early diagnosis of leukaemias and lymphomas(Cranfield University, 2013-11) Jackson, Olivia; Stone, Nicholas; Bessant, Conrad; Rye, Adam; Lush, Richard; McCarthy, KeithThis thesis aimed to investigate vibrational spectroscopies for the identification of biochemical markers of leukaemias and lymphomas. In a preliminary study using the blood proteins albumin, fibrinogen and globulin, Drop Coating Deposition Raman Spectroscopy was explored and extended for use with Fourier Transform infrared spectroscopy for leukaemia blood sample analysis. Due to low sample volumes and minimal preparation required it was identified as a potential alternative to blood centrifugation to obtain the buffy coat for analysis. These studies identified that it was capable of detecting low levels of protein from small, highly concentrated droplets. Thus this method, alongside cytospin centrifugation, was used for the spectroscopic analysis of different blood fractions. Due to the low number of lymphoma samples obtained, only a feasibility study is outlined in this thesis. Samples were collected from leukaemia patients and healthy volunteers. Infrared and Raman spectra were measured of whole blood and buffy coat samples cytospun onto slides and whole blood and plasma pipetted by drop coating deposition. Multivariate statistical analysis was employed to extract key spectral differences between the pathologies and develop classification models for diagnosing chronic lymphoblastic leukaemia from previously treated and untreated patient groups. Principal component analysis followed by linear discriminant analysis was employed to identify the largest variances in the data and leave one sample out cross validation evaluated the performance of the spectral models measured on different blood components in diagnosing leukaemia. The buffy coat infrared model correctly classified 59% of the spectra, and blood droplet Raman 62%. The treated and untreated groups were then combined, which improved classification to 83% for buffy coat infrared and 71% for blood droplet Raman. These findings highlight the potential of drop coating deposition spectroscopy of whole blood for leukaemia diagnosis, although further work is required to achieve a clinically validated method.