Cell Staining by Photo-activated Dye and Its Conjugate with Chitosan
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Photo-activated or “Caged” rhodamine dyes are the most useful for microscopic investigation of biological tissue by various fluorescent techniques. Novel precursor of the fluorescent dye (PFD813) has been studied for photosensitive staining of numerous animal cells. The functional rhodamine dye (Rho813) with intensive fluorescence has been obtained after photoactivation of its precursor PFD813 inside cells. The dye Rho813 has been successfully used for the optical detection of particular features in biological objects (HaCaT cells, HBL-100, MDCK, lymphocytes). Moreover, the chitosan conjugate with PFD molecules (“Chitosan-PFD813″) has been obtained and studied for the first time. The developed procedures and obtained data are important for further applications of novel precursors of fluorescent dyes (“caged” dyes) for microscopic probing of biological objects. As example, the synthesized “Chitosan-PFD813″ has been successfully applied in this study for intracellular transport visualization by fluorescent microscopy.
KeywordsCaged fluorescent dye Cell staining Photoactivation Biomembranes Lipids Conjugate Chitosan
Some parts of this work were supported by grant from Russian Scientific Foundation (Project 14-16-00046). We thank Dr. Svirshchevskaya E.V. and Ph.D.-student Generalov A.A. for cell cultivation and suggestions on cell staining experiments; Dr. Belov V. N. for preparation of the precursor of fluorescent dye.
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