Extraction of masked fluorescence peaks through synchronous fluorescence spectroscopy

Seema Devi; Meghdoot Mozumder; Nirmalya Ghosh; Asima Pradhan

doi:10.1117/12.907334

9 February 2012 Extraction of masked fluorescence peaks through synchronous fluorescence spectroscopy

Seema Devi, Meghdoot Mozumder, Nirmalya Ghosh, Asima Pradhan

Proceedings Volume 8225, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues X; 822517 (2012) https://doi.org/10.1117/12.907334
Event: SPIE BiOS, 2012, San Francisco, California, United States

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.

Citation Download Citation

Seema Devi, Meghdoot Mozumder, Nirmalya Ghosh, and Asima Pradhan "Extraction of masked fluorescence peaks through synchronous fluorescence spectroscopy", Proc. SPIE 8225, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues X, 822517 (9 February 2012); https://doi.org/10.1117/12.907334

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