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Nuclear Green™ Photo-Fixable DCS1 *5 mM DMSO Solution*

Live, apoptotic (Staurosporine treated cells), and dead (4% formaldehyde-treated) HeLa cells were stained with Nuclear Green™ Photo-Fixable DCS1 dye and imaged using a fluorescence microscope equipped with a FITC filter set. The images were captured before and after fixation with a 365 nm Transilluminator for 20 minutes.
Live, apoptotic (Staurosporine treated cells), and dead (4% formaldehyde-treated) HeLa cells were stained with Nuclear Green™ Photo-Fixable DCS1 dye and imaged using a fluorescence microscope equipped with a FITC filter set. The images were captured before and after fixation with a 365 nm Transilluminator for 20 minutes.
Live, apoptotic (Staurosporine treated cells), and dead (4% formaldehyde-treated) HeLa cells were stained with Nuclear Green™ Photo-Fixable DCS1 dye and imaged using a fluorescence microscope equipped with a FITC filter set. The images were captured before and after fixation with a 365 nm Transilluminator for 20 minutes.
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Physical properties
Molecular weight988.31
SolventDMSO
Spectral properties
Excitation (nm)502
Emission (nm)527
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
StorageFreeze (< -15 °C); Minimize light exposure
UNSPSC41116134

OverviewpdfSDSpdfProtocol


See also: Nucleus
Molecular weight
988.31
Excitation (nm)
502
Emission (nm)
527
Nuclear Green™ Photo-Fixable DCS1 is a unique fluorogenic, DNA-selective, and cell-impermeant dye for analyzing DNA content in dead, fixed, or apoptotic cells. It selectively stains cell membrane-compromised cells. It contains a DNA-binding fluorophore, thus readily binds to DNA upon interaction. Nuclear Green™ Photo-Fixable DCS1 has its green fluorescence significantly enhanced upon binding to DNA. In addition to its DNA-binding fluorophore, it contains a secondary DNA-photoreactive group. When needed, a moderate UV illumination (e.g., 350 nm) makes the DNA-bound dye readily react with DNA and makes the dye permanently conjugate with the DNA. It is optimized for surviving the cell fixation process better than common DNA dyes such as PI, DAPI, Hoechst, 7-AAD, DRAQ-5, DRAQ-7, or SYBR Green. It can be used in fluorescence imaging, microplate, and flow cytometry applications. This DNA-binding dye might be used for multicolor analysis of dead, fixed, or apoptotic cells with proper filter sets.

Platform


Fluorescence microscope

ExcitationFITC filter set
EmissionFITC filter set
Recommended plateBlack wall/clear bottom

Example protocol


AT A GLANCE

Spectral Properties

Ex/Em = 510/532 nm (bound to DNA)

SAMPLE EXPERIMENTAL PROTOCOL

Caution: The following protocol can be adapted for most cell types. Growth medium, cell density, the presence of other cell types, and factors may influence staining. Residual detergent on glassware may also affect the staining of many organisms and cause brightly stained material to appear in solutions with or without cells present.

  1. Add 2 to 10 µM of Nuclear Green™ Photo-Fixable DCS1 to fixed, dead, or apoptotic cells (whether in suspension or adherent) and incubate for 15 to 60 minutes.

    Note: To determine the optimal concentration for the desired result, it is advisable to test a wide range of dye concentrations in initial experiments.

    Optional: Wash the cells twice with Hanks and 20 mM HEPES buffer (HBSS) or a buffer of your choice. Then, fill the wells with fresh HBSS or growth medium.

  2. Observe the cells using a fluorescence microscope, fluorescence microplate reader, or flow cytometer equipped with the desired filter set. 

  3. Optional: Fix cells with 4% formaldehyde for 20 minutes at room temperature. Wash cells twice to get rid of any fixation solution.

    Optional: Photofix cells with moderate UV illumination at 365 nm.

Calculators


Common stock solution preparation

Table 1. Volume of DMSO needed to reconstitute specific mass of Nuclear Green™ Photo-Fixable DCS1 *5 mM DMSO Solution* to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.

0.1 mg0.5 mg1 mg5 mg10 mg
1 mM101.183 µL505.914 µL1.012 mL5.059 mL10.118 mL
5 mM20.237 µL101.183 µL202.366 µL1.012 mL2.024 mL
10 mM10.118 µL50.591 µL101.183 µL505.914 µL1.012 mL

Molarity calculator

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Spectrum


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spectrum

Spectral properties

Excitation (nm)502
Emission (nm)527

Images