Managò, Stefano (2016) A reliable Raman-spectroscopy-based approach for diagnosis, classification and follow-up of B-cell acute lymphoblastic leukemia. [Tesi di dottorato]

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Item Type: Tesi di dottorato
Lingua: English
Title: A reliable Raman-spectroscopy-based approach for diagnosis, classification and follow-up of B-cell acute lymphoblastic leukemia
Creators:
CreatorsEmail
Managò, Stefanos.manago@ibp.cnr.it
Date: 31 March 2016
Number of Pages: 126
Institution: Università degli Studi di Napoli Federico II
Department: Fisica
Scuola di dottorato: Ingegneria industriale
Dottorato: Tecnologie innovative per materiali, sensori ed imaging
Ciclo di dottorato: 28
Coordinatore del Corso di dottorato:
nomeemail
Cassinese, AntonioUNSPECIFIED
Tutor:
nomeemail
De Luca, Anna ChiaraUNSPECIFIED
Date: 31 March 2016
Number of Pages: 126
Uncontrolled Keywords: Raman spectroscopy, diagnosis, leukemia, blood cells
Settori scientifico-disciplinari del MIUR: Area 02 - Scienze fisiche > FIS/07 - Fisica applicata (a beni culturali, ambientali, biologia e medicina)
Date Deposited: 08 Apr 2016 13:45
Last Modified: 02 Nov 2016 13:41
URI: http://www.fedoa.unina.it/id/eprint/10938

Abstract

The aim of my PhD research project was to develop an advanced optical technique, based on Raman Spectroscopy (RS) and its integration with other optical (e.g. fluorescence microscopy, flow cytometry, Raman imaging) and biological modalities (e.g. Western Blotting), in order to provide a fast and reliable procedure for identification and classification of single cells (hemogram) from peripheral blood of healthy donors as well as of cancer B-ALL patients. The work can be divided into three main parts: • Deep study of Raman spectroscopy literature with particular attention to its sensitivity, specificity and bio-applications. • Development and characterization of the Raman microscope including the design of optics and development mathematical methods for analysis of the data (Principal Component Analysis). Since the aim of my thesis is to study living cells in vivo, with particular reference to normal and leukemia cells, the main challenges have been to find correct laser wavelength and intensity, alignment and calibration of the Raman system and finally its application to demonstrate its usefulness. • Completion of four experiments that demonstrate the suitability of Raman technique for studies of live normal and leukemia cells. Schematically, my research project concerns: (i) Raman identification and classification of three leukemia cell lines, that closely mimic the different differentiation/maturation stages of B-ALL cells; (ii) Spectroscopic regression assessment of leukemia after low- and not-cytotoxic dose treatments of methotrexate (MTX) and 6-mercaptopurine (6MP), two key drugs currently used in the B-ALL maintenance therapy; (iii) RS analysis of clinical samples collected from peripheral blood of patients with B-ALL; (iv) Raman identification and differentiation of the most important leukocyte populations isolated from peripheral blood of several human volunteers via conventional flow cytometry (granulocytes, monocytes, T cells, B cells and NK cells). The results presented here demonstrate that RS in conjunction with multivariate statistical technique has potential for rapid label-free diagnosis, classification and follow-up after chemotherapy treatment of B-ALL based on the optical evaluation of spectral features of biomolecules.

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