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Conjugates of a photoactivated rhodamine with biopolymers for cell staining.

Zaitsev SY, Shaposhnikov MN, Solovyeva DO, Solovyeva VV, Rizvanov AA - ScientificWorldJournal (2014)

Bottom Line: In this study, a precursor of the photoactivated fluorescent dye (PFD) has been successfully used for staining of numerous mammalian cells lines and for conjugate formation with chitosan ("Chitosan-PFD") and histone H1 ("Histone H1.3-PFD").The intensive fluorescence has been observed after photoactivation of these conjugates inside cells (A431, HaCaT, HEK239, HBL-100, and MDCK).Developed procedures and obtained data are important for further application of novel precursors of fluorescent dyes ("caged" dyes) for microscopic probing of biological objects.

View Article: PubMed Central - PubMed

Affiliation: Moscow State Academy of Veterinary Medicine and Biotechnology, Akad. Skryabin Street 23, Moscow 109472, Russia.

ABSTRACT
Conjugates of the photoactivated rhodamine dyes with biopolymers (proteins, polysaccharides, and nucleic acids) are important tools for microscopic investigation of biological tissue. In this study, a precursor of the photoactivated fluorescent dye (PFD) has been successfully used for staining of numerous mammalian cells lines and for conjugate formation with chitosan ("Chitosan-PFD") and histone H1 ("Histone H1.3-PFD"). The intensive fluorescence has been observed after photoactivation of these conjugates inside cells (A431, HaCaT, HEK239, HBL-100, and MDCK). Developed procedures and obtained data are important for further application of novel precursors of fluorescent dyes ("caged" dyes) for microscopic probing of biological objects. Thus, the synthesized "Chitosan-PFD" and "Histone H1-PFD" have been successfully applied in this study for intracellular transport visualization by fluorescent microscopy.

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HaCaT cells stained by conjugate “Chitosan-PFD” (conc. 2.2 mg/mL) (a), the mixture of PFD (conc. 5 μg/mL) and chitosan (2.2 μg/mL) (b), and PFD-NHS (conc. 5 μg/mL) and chitosan (2.2 μg/mL) (c) after photoactivation (image dimensions, 36 by 36 μm).
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fig4: HaCaT cells stained by conjugate “Chitosan-PFD” (conc. 2.2 mg/mL) (a), the mixture of PFD (conc. 5 μg/mL) and chitosan (2.2 μg/mL) (b), and PFD-NHS (conc. 5 μg/mL) and chitosan (2.2 μg/mL) (c) after photoactivation (image dimensions, 36 by 36 μm).

Mentions: The rhodamine derivatives can be used to study biopolymer transport using fluorescent microscopy. Chitosan is important biopolymer for modeling and applied studies of the BAS transport in various cells. Chitosan heterogeneous clusters in the cells were observed around the nucleus after photoactivation (Figure 4(a)). The images of cells stained with the PFD (or PFD-813-NHS) in the presence of chitosan are shown in Figures 4(b) and 4(c) for comparison. In the case of last two mixtures, dye distribution inside cell was more uniform (Figures 4(b) and 4(c)) as compared to the conjugate (Figure 4(a)). This can be a consequence of the free dye distribution (separately from the chitosan) in the case of the abovementioned mixtures. Moreover, the synthesized conjugate of chitosan with PFD molecules (“Chitosan-PFD”) is suitable for chitosan imaging inside the cells using laser scanning confocal microscopy. The discussed conjugate is just one example of such PFD application in the study of molecular transport visualized by fluorescent microscopy.


Conjugates of a photoactivated rhodamine with biopolymers for cell staining.

Zaitsev SY, Shaposhnikov MN, Solovyeva DO, Solovyeva VV, Rizvanov AA - ScientificWorldJournal (2014)

HaCaT cells stained by conjugate “Chitosan-PFD” (conc. 2.2 mg/mL) (a), the mixture of PFD (conc. 5 μg/mL) and chitosan (2.2 μg/mL) (b), and PFD-NHS (conc. 5 μg/mL) and chitosan (2.2 μg/mL) (c) after photoactivation (image dimensions, 36 by 36 μm).
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4214035&req=5

fig4: HaCaT cells stained by conjugate “Chitosan-PFD” (conc. 2.2 mg/mL) (a), the mixture of PFD (conc. 5 μg/mL) and chitosan (2.2 μg/mL) (b), and PFD-NHS (conc. 5 μg/mL) and chitosan (2.2 μg/mL) (c) after photoactivation (image dimensions, 36 by 36 μm).
Mentions: The rhodamine derivatives can be used to study biopolymer transport using fluorescent microscopy. Chitosan is important biopolymer for modeling and applied studies of the BAS transport in various cells. Chitosan heterogeneous clusters in the cells were observed around the nucleus after photoactivation (Figure 4(a)). The images of cells stained with the PFD (or PFD-813-NHS) in the presence of chitosan are shown in Figures 4(b) and 4(c) for comparison. In the case of last two mixtures, dye distribution inside cell was more uniform (Figures 4(b) and 4(c)) as compared to the conjugate (Figure 4(a)). This can be a consequence of the free dye distribution (separately from the chitosan) in the case of the abovementioned mixtures. Moreover, the synthesized conjugate of chitosan with PFD molecules (“Chitosan-PFD”) is suitable for chitosan imaging inside the cells using laser scanning confocal microscopy. The discussed conjugate is just one example of such PFD application in the study of molecular transport visualized by fluorescent microscopy.

Bottom Line: In this study, a precursor of the photoactivated fluorescent dye (PFD) has been successfully used for staining of numerous mammalian cells lines and for conjugate formation with chitosan ("Chitosan-PFD") and histone H1 ("Histone H1.3-PFD").The intensive fluorescence has been observed after photoactivation of these conjugates inside cells (A431, HaCaT, HEK239, HBL-100, and MDCK).Developed procedures and obtained data are important for further application of novel precursors of fluorescent dyes ("caged" dyes) for microscopic probing of biological objects.

View Article: PubMed Central - PubMed

Affiliation: Moscow State Academy of Veterinary Medicine and Biotechnology, Akad. Skryabin Street 23, Moscow 109472, Russia.

ABSTRACT
Conjugates of the photoactivated rhodamine dyes with biopolymers (proteins, polysaccharides, and nucleic acids) are important tools for microscopic investigation of biological tissue. In this study, a precursor of the photoactivated fluorescent dye (PFD) has been successfully used for staining of numerous mammalian cells lines and for conjugate formation with chitosan ("Chitosan-PFD") and histone H1 ("Histone H1.3-PFD"). The intensive fluorescence has been observed after photoactivation of these conjugates inside cells (A431, HaCaT, HEK239, HBL-100, and MDCK). Developed procedures and obtained data are important for further application of novel precursors of fluorescent dyes ("caged" dyes) for microscopic probing of biological objects. Thus, the synthesized "Chitosan-PFD" and "Histone H1-PFD" have been successfully applied in this study for intracellular transport visualization by fluorescent microscopy.

Show MeSH