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Proceedings Paper

Extraction of masked fluorescence peaks through synchronous fluorescence spectroscopy
Author(s): Seema Devi; Meghdoot Mozumder; Nirmalya Ghosh; Asima Pradhan
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Paper Abstract

Fluorescence spectroscopy has been demonstrated as a viable tool for noting subtle biochemical changes that occur during early-stage cervical cancer progression. Due to multiple fluorophore contributions, the individual fluorophore activities often get masked due to overlapping spectra of neighboring fluorophores. Recently synchronous fluorescence spectroscopy has been demonstrated as an efficient technique for investigation of such non-dominant fluorophores. With synchronous fluorescence spectroscopy individual fluorophore responses are highlighted as sharp peaks by choosing appropriate offsets during signal acquisition. Such peaks may, however be missed due to absorption effects. By correcting the measured synchronous fluorescence spectrum with elastic scattering data, it was observed that the masked fluorophores are highlighted while the broader bands are sharpened. Interestingly, fluorophore activities of protoporphyrin, collagen, NADH, FAD and porphyrin can now be studied using this technique, as compared to only collagen and NADH seen earlier. The results have been verified using tissue phantoms with known fluorophores and scatterers. Use of normalized synchronous spectra has led to enhancement of several fluorophore responses. It was also observed that among the different offsets, the lower ones show sharper features, whereas the larger offsets show a broadband response. Among the different offsets 120nm is found optimal for further investigation.

Paper Details

Date Published: 9 February 2012
PDF: 6 pages
Proc. SPIE 8225, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues X, 822517 (9 February 2012); doi: 10.1117/12.907334
Show Author Affiliations
Seema Devi, Indian Institute of Technology Kanpur (India)
Meghdoot Mozumder, Indian Institute of Technology Kanpur (India)
Nirmalya Ghosh, Indian Institute of Science Education and Research Kolkata (India)
Asima Pradhan, Indian Institute of Technology Kanpur (India)


Published in SPIE Proceedings Vol. 8225:
Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues X
Daniel L. Farkas; Dan V. Nicolau; Robert C. Leif, Editor(s)

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