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Multicolor Labeling of Cell Nucleus with DNA-Binding Dyes

The nucleus is the largest cellular organelle in animals. In mammalian cells, the average diameter of the nucleus is approximately 6 µm, which occupies about 10% of the total cell volume. Nucleus contains most of the cell's genetic material, organized as multiple long linear DNA molecules in complex with a large variety of proteins, such as histones, to form chromosomes. The genes within these chromosomes are the cell's nuclear genome. The function of the nucleus is to maintain the integrity of these genes and to control the activities of the cell by regulating gene expression, therefore, the nucleus is the control center of the cell. The main structures making up the nucleus are the nuclear envelope, a double membrane that encloses the entire organelle and isolates its contents from the cellular cytoplasm, and the nucleoskeleton. Movement of large molecules through the pores is required for both gene expression and the maintenance of chromosomes.

 

Labeling the Nuclei of Live Cells



Image of live HeLa cells stained with Nuclear Green™ LCS1. The mitochondria of live HeLa cells were stained with red fluorescence Cell Navigator® Mitochondrion Staining Kit.
Nuclear Green™ LCS1, Nuclear Orange™ LCS1, Nuclear Red™ LCS1 and Nuclear Yellow are fluorogenic, DNA-selective and cell-permeant dyes for analyzing DNA content in living cells. The fluorescence of these dyes is significantly enhanced upon binding to DNA. They can be used in fluorescence imaging, microplate and flow cytometry applications. These DNA-binding dyes might be used for multicolor analysis of live cells with proper filter sets.

Our recently developed Nuclear Blue™ LCS1 is a fluorogenic, DNA-selective and cell-permeant dye for analyzing DNA content in living cells. The Nuclear Blue™ LCS1 has its blue fluorescence significantly enhanced upon binding to DNA. It can be used in fluorescence imaging, microplate and flow cytometry applications. It is well excited by violet laser at 405 nm, and emits blue/cyan fluorescence light with an emission maximum at ~440 nm, and provides an excellent tool for flow cytometers equiped with a 405 nm violet laser source. This DNA-binding dye might be used for multicolor analysis of live cells with the filter sets of Pacific Blue™ and BD Horizon™ V450.



Labeling the Nuclei of Dead Cells



Binding activity of Apopxin™ Deep Red to phosphatidylserine in Jurkat cells. The fluorescence images demonstrated that live cells (blue) were stained by CytoCalcein™ Violet 450, apoptotic cells (red) were stained by Apopxin™ Deep Red, and necrotic cells (green) were stained by Nuclear Green™ DCS1. Apoptosis was induced by 1 µM staurosporine for 3 hours. The fluorescence images of the cells were taken with Olympus fluorescence microscope using the violet, Cy5® and FITC channel respectively. A: Non-induced control cells; B: Triple staining of staurosporine-induced cells.

The normalized emission spectral comparison of Nuclear Green™ DCS1 (Ex/Em = 503/526 nm), Nuclear Orange™ DCS1 (Ex/Em = 514/555 nm), and Nuclear Red™ DCS1 (Ex/Em = 622/645 nm) in the presence of calf thymus DNA. The dotted line is emission spectrum of propidium iodide bound to DNA (Ex/Em = 535/617 nm).
Nuclear Green™ DCS1, Nuclear Orange™ DCS1 and Nuclear Red™ DCS1 are fluorogenic, DNA-selective and cell-impermeant dyes for analyzing DNA content in dead, fixed or apoptotic cells. As the LCS1 reagents, the fluorescence of the DCS1 dyes is significantly enhanced upon binding to DNA. They can be used in fluorescence imaging, microplate and flow cytometry applications. These DNA-binding dyes might be used for multicolor analysis of dead, fixed or apoptotic cells with proper filter sets.



















Table 1. Cell Nuclear Stains.
Cat# Product Name Ex (nm) Em (nm) Size Price
1300 ReadiLink™ Rapid trFluor™ Eu Antibody Labeling Kit *Microscale Optimized for Labeling 50 ug Antibody Per Reaction* 346 617 2 Labelings $950
17510 DAPI [4,6-Diamidino-2-phenylindole, dihydrochloride] *CAS 28718-90-3* 358 461 10 mg $75
17562 Hexidium Iodide *CAS 211566-66-4* 518 600 5 mg $145
17520 Hoechst 33258 *CAS 23491-45-4* 352 461 100 mg $95
17530 Hoechst 33342 *Ultrapure Grade* *CAS 23491-52-3* 350 461 100 mg $95
17537 Hoechst 34580 *CAS 911004-45-0* 368 437 5 mg $145
17514 Hydroxystilbamidine *CAS 223769-64-0* 360 591 10 mg $95
17561 LDS 751 *CAS 181885-68-7* 543 712 25 mg $95
17543 Nuclear Violet™ LCS1 401 459 0.5 ml $95
17550 Nuclear Green™ DCS1 503 526 0.5 ml $95
17540 Nuclear Green™ LCS1 503 526 0.5 ml $95
17551 Nuclear Orange™ DCS1 528 576 0.5 ml $95
17541 Nuclear Orange™ LCS1 514 555 0.5 ml $95
17552 Nuclear Red™ DCS1 642 660 0.5 ml $95
17542 Nuclear Red™ LCS1 622 645 0.5 ml $95
17539 Nuclear Yellow [Hoechst S769121] *CAS 74681-68-8* 355 495 25 mg $195
17515 Propidium iodide *CAS 25535-16-4* 535 617 25 mg $95