Discrimination of normal and cancerous human skin tissues based on laser-induced spectral shift fluorescence microscopy.

A homemade spectral shift fluorescence microscope (SSFM) is coupled with a spectrometer to record the spectral images of specimens based on the emission wavelength. Here a reliable diagnosis of neoplasia is achieved according to the spectral fluorescence properties of ex-vivo skin tissues after rhodamine6G (Rd6G) staining. It is shown that certain spectral shifts occur for nonmelanoma/melanoma lesions against normal/benign nevus, leading to spectral micrographs. In fact, there is a strong correlation between the emission wavelength and the sort of skin lesions, mainly due to the Rd6G interaction with the mitochondria of cancerous cells. The normal tissues generally enjoy a significant red shift regarding the laser line (37 nm). Conversely, plenty of fluorophores are conjugated to unhealthy cells giving rise to a relative blue shift i.e., typically SCC (6 nm), BCC (14 nm), and melanoma (19 nm) against healthy tissues. In other words, the redshift takes place with respect to the excitation wavelength i.e., melanoma (18 nm), BCC (23 nm), and SCC (31 nm) with respect to the laser line. Consequently, three data sets are available in the form of micrographs, addressing pixel-by-pixel signal intensity, emission wavelength, and fluorophore concentration of specimens for prompt diagnosis.

Fluorescence emission quenching of RdB fluorophores in attendance of various blood type RBCs based on Stern-Volmer formalism.

In this work, the optical properties of Rhodamine B (RdB) are investigated in the attendance of various red blood cells (RBCs). RdB fluorophores, as biological markers, is excited using SHG-CW Nd:YAG laser at 532 nm. In fact, the addition of biomolecules of interest to the reference fluorophore notably changes the fluorescence properties of the suspension. Here, laser induced fluorescence (LIF) spectrophotometry based on Stern-Volmer quenching formalism and field emission scanning electron microscope (FESEM) are employed here. According to the given fluorescence spectra, the spectral shift of emissions as well as quenching coefficients are assessed subsequently. The Stern-Volmer formalism is used to determine the quenching coefficients. In fact, RdB + RBCs suspensions contain a plenty of bioconjugates leading to the signal reduction and notable red shift in RdB fluorescence emissions. Furthermore, it is demonstrated that the positive blood type RBCs exhibit the higher quenching coefficients and the larger red shifts against those of negative blood types. This mainly arises from the nature of specific sugar antigens available on the RBC membranes as to N-acetylgalactosamine and galactose attached to the O-antigen terminal would enhance further quenching of the species. Moreover, a significant correlation appears between Stern-Volmer coefficients and the corresponding RBCs. In fact, distinct discrepancy takes place in quenching coefficients in terms various positive/negative blood types to envisage a facile method of blood typing.

Fluorescence properties of Phycocyanin and Phycocyanin-human serum albumin complex.

In this work, the fluorescence properties of Phycocyanin (PC) and the corresponding quenching effects are investigated in attendance of human serum albumin (HSA). At first, PC is excited at 532 nm using CW SHG Nd:YAG laser, then the emission wavelength, Stokes shift, quantum yield, extinction constant and self-quenching coefficient are obtained based on the modified Beer-Lambert equation. It is shown that a notable red shift appears in terms of PC concentration. According to the fluorescence spectra, the addition of HSA in PC solution leads to a significant reduction in the fluorescence signal via quenching events, however a lucid blue shift takes place in the same time. Stern-Volmer formalism is used to determine the quenching constant (KS), the number of binding sites (n) between PC and HSA as well as the association constant Ka for the purpose of facile transportation to the target in the context of drug delivery. Eventually, temperature dependent coefficients and corresponding spectral shifts are investigated over a wide range of temperatures at a couple of distinct PC concentrations to attest the dominant static quenching takes place. The rate of conjugate formations elevates at low temperatures leading to a certain blue shift. Furthermore, large KS is measured in the course of signal reduction, particularly at low PC populations. In fact, PC conjugation to HSA is essential interaction to enhance chemo drug transportation. Here, at the body temperature, the quenching coefficient decreases to facilitate the drug release. Moreover, the spectral shift of fluorescence emission can be useful for simultaneous monitoring and drug delivery treatment.