Fluorescence labelling is one of the significant properties for optical bioimaging and biosensing applications. The development in the field of nanotechnology has created enormous conveniences in biomedical research. The fluorescence enhancement of mammalian cell in presence of zinc nitride colloidal nanoparticle was imaged using multiphoton confocal microscope. The statistical significance in the presence and absence of nanoparticle evidenced the fluorescence intensity enhancement in the cells. In order to suppress the background signal and increase the penetration depth, the multiphoton technique is utilized by 740 nm excitation to examine the enhancement potential of Zn3N2 colloidal nanoparticles towards cell structures. Hence, we suggest that, the unique feature of inorganic nanomaterial is a promising material for probing cells for future diagnostic applications.
Metabolic imaging of live cell may allow in understanding the molecular level changes in cells under various diseased state, including cancer. The intrinsic fluorophores, Nicotinamide adenine dinucleotide (NADH) and flavin adenine dinucleotide (FAD) are crucial for electron transfer in the oxidation-reduction reactions in the cell. Metabolic imaging based on fluorescence polarization enables to analyze both biochemical distribution and their conformation. In this study, multiphoton fluorescence polarization imaging of NADH and FAD from cancer and normal cell lines of epithelial origin were carried out. Spectral deconvolution method was adopted to isolate fluorescence emission from different coenzymes NADH and FAD. The observed heterogeneity of the multiphoton autofluorescence in live cells was used in intensity-toconcentration image conversion. The multiphoton autofluorescence exhibits anisotropy features at the cellular level, that directly indicate the presence of NADH and FAD in two differing conformation states viz; free and protein-bound. Mapping of anisotropy of cellular autofluorescence enables to probe the distribution of population fractions of free and bound forms of NADH and FAD. Further, the redox ratio between normal and cancer cell lines confirms the changes in the metabolic activities between them. These molecular-level studies demonstrate the potential of probing cellular metabolism associated with cancer, without the need for cell destruction as in the case of conventional biochemical assays.
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