![]() ![]() 5 Principal component analysis (PCA) 6, 7 and two-dimensional correlation spectroscopy (2D-COS) and its variants 8– 12 are dependent on the analysis of correlated changes between variables. Nevertheless, a data search for those features that are linearly the most dissimilar to each other often becomes necessary because reduced complexity is desirable to implement many approaches such as multivariate curve resolution (MCR). Though Raman marker bands, unique to a given component, can be useful for identification and quantification purposes, 4 this is often not possible when substantial band overlapping occurs. To identify the individual components and their relative concentrations, there is a need to “demix” such superpositions. The Raman spectra of biological cell and tissue samples consist of the superposition of scattering from numerous different macromolecular components. Our efforts provided insight into the need for effective resolution enhancement approaches, the feasibility of these methods for automation, the nature of the problems currently limiting their use, and in particular those aspects that need improvement. We conclude with a discussion of the particular benefits, drawbacks, and potential of these methods. ![]() We also determined the effects of the different resolution enhancement methods on 2D-COS and on chemical contrast image generation from mammalian cell spectra. High resolution spectra of glucose acquired with a narrow spectrometer slit were compared to ones using a wide slit that degraded the spectral resolution. We then investigated the efficacy of these methods to improve the resolution of measured Raman spectra. Improved spectral resolution was evaluated based on several criteria including the separation of overlapping peaks and the preservation of true peak intensities in resolution-enhanced spectra. We first applied the methods to varied synthetic spectra, each consisting of nine overlapping Voigt profile peaks. To investigate these issues, we implemented three novel resolution enhancement methods based on pseudospectra, over-deconvolution, and peak fitting to evaluate them along with three extant methods: node narrowing, blind deconvolution, and the general-purpose peak fitting program Fityk. Furthermore, there is a need for automated methods to apply to large numbers of varied hyperspectral data sets containing multiple overlapping peaks, and thus methods ideally suitable for diverse tasks. Though software-based resolution enhancement methods can be used to counter such problems, their performances often differ, thereby rendering some more suitable than others for specific tasks. This is particularly problematic for methods dependent on sparsity such as multivariate curve resolution and other spectral demixing as well as for two-dimensional correlation spectroscopy (2D-COS), multisource correlation analysis, and principal component analysis. Overlapping peaks in Raman spectra complicate the presentation, interpretation, and analyses of complex samples. All subjects Allied Health Cardiology & Cardiovascular Medicine Dentistry Emergency Medicine & Critical Care Endocrinology & Metabolism Environmental Science General Medicine Geriatrics Infectious Diseases Medico-legal Neurology Nursing Nutrition Obstetrics & Gynecology Oncology Orthopaedics & Sports Medicine Otolaryngology Palliative Medicine & Chronic Care Pediatrics Pharmacology & Toxicology Psychiatry & Psychology Public Health Pulmonary & Respiratory Medicine Radiology Research Methods & Evaluation Rheumatology Surgery Tropical Medicine Veterinary Medicine Cell Biology Clinical Biochemistry Environmental Science Life Sciences Neuroscience Pharmacology & Toxicology Biomedical Engineering Engineering & Computing Environmental Engineering Materials Science Anthropology & Archaeology Communication & Media Studies Criminology & Criminal Justice Cultural Studies Economics & Development Education Environmental Studies Ethnic Studies Family Studies Gender Studies Geography Gerontology & Aging Group Studies History Information Science Interpersonal Violence Language & Linguistics Law Management & Organization Studies Marketing & Hospitality Music Peace Studies & Conflict Resolution Philosophy Politics & International Relations Psychoanalysis Psychology & Counseling Public Administration Regional Studies Religion Research Methods & Evaluation Science & Society Studies Social Work & Social Policy Sociology Special Education Urban Studies & Planning BROWSE JOURNALS ![]()
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