Methods and instrumentation for raman characterization of bladder cancer tumor

High incidence and recurrence rates make bladder cancer the most common malignant tumor in the urinary system. Cystoscopy is the gold standard test used for diagnosis, nevertheless small flat tumors might be missed, and the procedure still represents discomfort to patients and high recurrence can result from of urethral injuries. During cystoscopy, suspicious tumors are detected through white light endoscopy and resected tissue is further examined by histopathology. after resection, the pathologist provides information on the differentiation of the cells and the penetration depth of the tumor in the tissue, known as grading and staging of tumor, respectively. During cystoscopy, information on tumor grading and morphological depth characterization can assist onsite diagnosis and significantly reduce the amount of unnecessarily resected tissue. Recently, new developments in optical imaging and spectroscopic approaches have been demonstrated to improve the results of standard techniques by providing real-time detection of macroscopic and microscopic biomedical information. Different applications to detect anomalies in tissues and cells based on the chemical composition and structure at the microscopic level have been successfully tested. There is, nevertheless, the need to cope with the demands for clinical translation. This doctoral thesis presents the investigations, clinical studies and approaches applied to filling the main open research questions when applying Raman spectroscopy as a diagnostic tool for bladder cancer tumor grading and general Raman spectroscopy-based oncological clinical studies

In-vivo Raman spectroscopy: from basics to applications

For more than two decades, Raman spectroscopy has found widespread use in biological and medical applications. The instrumentation and the statistical evaluation procedures have matured, enabling the lengthy transition from ex-vivo demonstration to in-vivo examinations. This transition goes hand-in-hand with many technological developments and tightly bound requirements for a successful implementation in a clinical environment, which are often difficult to assess for novice scientists in the field. This review outlines the required instrumentation and instrumentation parameters, designs, and developments of fiber optic probes for the in-vivo applications in a clinical setting. It aims at providing an overview of contemporary technology and clinical trials and attempts to identify future developments necessary to bring the emerging technology to the clinical end users. A comprehensive overview of in-vivo applications of fiber optic Raman probes to characterize different tissue and disease types is also given

Using Raman Spectroscopy for Intraoperative Margin Analysis in Breast Conserving Surgery

Breast Conserving Surgery (BCS) in the treatment of breast cancer aims to provide optimal oncological results, with minimal tissue excision to optimise cosmetic outcome. Positive margins due to an inadequate resection occurs in 17% of UK patients undergoing BCS and prompts recommendation for further tissue re-excision to reduce recurrence risk. A second operation causes patient anxiety and significant healthcare costs. This issue could be resolved with accurate intra-operative margin analysis (IMA) to enable excision of all cancerous tissue at the index procedure. High wavenumber Raman Spectroscopy (HWN RS) is a vibrational spectroscopy highly sensitive to changes in protein/lipid environment and water content –biochemical differences found between tumour and normal breast tissue. We proposed that HWN RS could be used to differentiate between tumour and non-tumour breast tissue with a view to future IMA. This thesis presents the development of a Raman system to measure the HWN region capable of accurately detecting changes in protein, lipid and water content, in the presence of highly fluorescent surgical pigments such as blue dye that are present in surgically excised specimens. We investigate the relationship between changes in the HWN spectra with changes in water content in constructed breast phantoms to mimic protein and lipid rich environments and biological tissue. Human breast tissue of paired tumour and non-tumour samples were then measured and analysed. We found that breast tumour tissue is a protein rich, high water, low fat environment and that non-tumour is a low protein, fat rich environment with a low water content, and this can be used to identify breast cancer using HWN RS with excellent accuracy of over 90%. This thesis demonstrates a HWN RS Raman system capable of differentiating between tumour and non-tumour tissue in human breast tissue, and this has the potential to provide IMA in BCS

Spatially offset Raman spectroscopy

This is the author accepted manuscript. The final version is available from Springer Nature via the DOI in this recordSpatially offset Raman spectroscopy (SORS) is a spectroscopic technique that allows for the non-invasive chemical characterization of diffusely scattering materials, ranging from opaque plastics to biological tissues. SORS has been explored for a range of applications, including disease diagnosis, the detection of explosives through unopened containers and the in-depth, non-destructive analysis of pharmaceutical products and objects of art. This Primer introduces the reader to the basic concepts underpinning SORS, details best practices for its implementation, highlights its use across multiple fields and provides insight into its limitations. The Primer concludes by discussing potential applications and envisaging future developments in the field.Engineering and Physical Sciences Research Council (EPSRC

Near-infrared raman spectroscopy for early detection of cervical precancer

Ph.DDOCTOR OF PHILOSOPH

Improving Clinical Diagnosis of Melanocytic Skin Lesions by Raman Spectroscopy

High-quality Raman signals from melanocytic lesions compatible with a possible clinical application have not been demonstrated yet. The objectives of the work described in this thesis were: I: The development of a Raman spectroscopic prototype for objective and fast assessment of melanocytic skin lesions clinically suspicious for melanoma; II: Identification of the main spectroscopic features of melanoma and benign melanocytic lesions suspicious for melanoma; III: Assessment of the feasibility of Raman spectroscopy as an adjunct technique to improve clinical diagnosis of melanocytic skin lesions

Near-Infrared Confocal Raman Spectroscopy for Real-Time Diagnosis of Cervical Precancer

Ph.DDOCTOR OF PHILOSOPH

Recent Developments in Atomic Force Microscopy and Raman Spectroscopy for Materials Characterization

This book contains chapters that describe advanced atomic force microscopy (AFM) modes and Raman spectroscopy. It also provides an in-depth understanding of advanced AFM modes and Raman spectroscopy for characterizing various materials. This volume is a useful resource for a wide range of readers, including scientists, engineers, graduate students, postdoctoral fellows, and scientific professionals working in specialized fields such as AFM, photovoltaics, 2D materials, carbon nanotubes, nanomaterials, and Raman spectroscopy

Raman spectroscopy for unlabelled detection and quantification of drugs in tissue

Raman spectroscopy is an optical analysis technique for chemical analysis of samples. Inthis thesis, we have assessed its applicability in quantitative detection of drugs within animal tissue. This involved the design, construction, and optimisation of a Raman microscope for this application, considering the specific requirements of the samples. A Raman microscope was designed to efficiently measure Raman spectra from thin (16 μm) tissue sections with >1 cm field-of-view. Two of these were built, one with a 671 nm and one with a 785 nm wavelength excitation laser, to assess the relative benefits of each for drug detection specifically in the Raman silent region of the Raman spectrum (~ 1800 – 2800 cm-1), in both low and highly autofluorescing tissue. These instruments, and the acquisition parameters used, were optimised to maximise Raman throughput while minimising the effects of noise on the measurements from autofluorescence. Control tissue cryosections with drug solution pipetted on top was used as a feasibility test for qualitative detection of drugs in tissue. A model for prediction of the spectra and required measurement times for detection of different drug/tissue combinations at different concentrations was developed. The most promising drugs from this study were then used to generate mimetic tissue models, homogenous mixtures of drug and tissue with known mass ratios, to assess the quantifiability of Raman spectroscopy in drug detection in tissue. Detection limits as low as 18 μg/g were calculated for ponatinib in rat brain mimetic tissue models with 2-hour measurement times, and 34 μg/g for the ponatinib in rat liver mimetic tissue models

REAL-TIME FIBER OPTIC RAMAN SPECTROSCOPY FOR EARLY DIAGNOSIS OF PRECANCER AND CANCER IN VIVO IN THE UPPER GASTROINTESTINAL TRACT

Ph.DDOCTOR OF PHILOSOPH