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• W2012157299 startingPage "147" @default.
• W2012157299 abstract "The objective of this study was to implement a simple and reliable fluorescent staining method specific to DNA for the visualization of cell nuclei using 488-nm and 543-nm visible lasers or conventional mercury lamps. The staining properties of two different reagents, chromomycin A, (C-A3) and 7-aminoactinomycin-D (7-AA-D), were analyzed in several types of mammalian cells and tissues with different fixation and preparation methods. Results were obtained using a scanning laser confocal microscope. Experiments were initially conducted in paraformaldehyde-fixed cell cultures to optimize the staining conditions. C-A3 (100 μg/mL) and 7-AA-D (10 μM) showed consistent and specific binding to nuclear DNA and emitted green (-580nm) and red (-647nm) colors, respectively. No dose change was needed for formaldehyde-fixed cells in the same conditions. In paraffin-embedded or frozen sections, C-A3 exhibited optimum labeling at 140 μglmL, whereas 7-AA-D was found optimum at 10 μM concentration. Both dyes displayed similar staining patterns regarding the specificity to chromatin and chromosomes but not to the RNA molecules. Major RNA-containing regions in cells, such as nucleolus or cytoplasm, were entirely devoid of C-A3 and 7-AA-D staining. All stages of mitotic chromosomes, as well as picnotic and apoptotic cells, were successfully labeled. Penetration of dyes for labeling giant cells (e.g., mouse oocytes) was achieved by a mild detergent extraction procedure. C-A3 and 7-AA-D are highly specific, easily handled fluorescent markers in PFA and FA fixed cultured cells, paraffin-embedded, and frozen sections. They can easily be the first choice for a specific DNA labeling as a primary or a counterstain in any fluorescent microscope, especially in confocal microscopes lacking UV laser. (The J Histotechnol 26:147, 2003)Submitted April 17, 2003; Accepted July 31, 2003" @default.
• W2012157299 created "2016-06-24" @default.
• W2012157299 creator A5062778222 @default.
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• W2012157299 date "2003-09-01" @default.
• W2012157299 modified "2023-09-26" @default.
• W2012157299 title "Two Convenient Methods for Nuclear Labeling in Confocal Microscopy using Visible-Lasers" @default.
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• W2012157299 doi "https://doi.org/10.1179/his.2003.26.3.147" @default.
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