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DiYO™-3 [equivalent to YOYO®-3] *5 mM DMSO Solution* *CAS 156312-20-8*

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Physical properties
Molecular weight1322.73
Spectral properties
Extinction coefficient (cm -1 M -1)1670001
Excitation (nm)612
Emission (nm)631
Quantum yield0.151
Storage, safety and handling
H-phraseH303, H313, H340
Hazard symbolT
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R68
StorageFreeze (< -15 °C); Minimize light exposure


Molecular weight
Extinction coefficient (cm -1 M -1)
Excitation (nm)
Emission (nm)
Quantum yield
DiYO™-3 is chemically equivalent to YOYO®-3 (YOYO® is the trademark of Invitrogen). DiYO™-3 is a carbocyanine dimer with far-red fluorescence similar to Cy® 5 dyes. It is cell-impermeant and easily distinguished from fluorescein and rhodamine as a nuclear counterstain and dead cell indicator. It is non-fluorescent in the absence of nucleic acids, and generates a very bright fluorescence signal upon binding to DNA. DiYO™-3 gives strong and selective nuclear staining in cultured cells and in paraffin sections. Simultaneous labeling with cell-permeable Nuclear Green™ LCS1 dye and cell-impermeant DiYO®-3 can be used to assess cell viability. DiYO™-3 and Nuclear Green™ both have much greater extinction coefficients than that of DNA-bound propidium iodide.


Fluorescence microscope

ExcitationCy5 filter set
EmissionCy5 filter set
Recommended plateBlack wall/clear bottom
Instrument specification(s)Cy5 filter set

Example protocol


DiYO™-3 working solution
Make DiYO™-3 working solution in Hanks with 20 mM Hepes buffer (HH buffer) or buffer of your choice at your desired concentration.
Note     In initial experiments, it may be best to try several dye concentrations to determine the optimal concentration that yields the desired result. High dye concentration tends to cause nonspecific staining of other cellular structures.


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 staining of many organisms, and cause brightly stained material to appear in solutions with or without cells present.
  1. Grow and treat cells as desired.
  2. Remove the cell culture medium and fix cells.
  3. Add DiYO™-3 working solution (1 to 10 µM) into the cells (either suspension or adherent cells), and stain the cells for 15 to 60 minutes.
  4. Remove the dye working solution and add HH buffer or buffer of your choice.
  5. Analyze the cellular staining with a fluorescence microscope using Cy5 filter. 


Common stock solution preparation

Table 1. Volume of DMSO needed to reconstitute specific mass of DiYO™-3 [equivalent to YOYO®-3] *5 mM DMSO Solution* *CAS 156312-20-8* 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 mM75.601 µL378.006 µL756.012 µL3.78 mL7.56 mL
5 mM15.12 µL75.601 µL151.202 µL756.012 µL1.512 mL
10 mM7.56 µL37.801 µL75.601 µL378.006 µL756.012 µL

Molarity calculator

Enter any two values (mass, volume, concentration) to calculate the third.

Mass (Calculate)Molecular weightVolume (Calculate)Concentration (Calculate)Moles


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Spectral properties

Extinction coefficient (cm -1 M -1)1670001
Excitation (nm)612
Emission (nm)631
Quantum yield0.151



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Authors: Cosa G, Focsaneanu KS, McLean JR, McNamee JP, Scaiano JC.
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TO-PRO-3 is an optimal fluorescent dye for nuclear counterstaining in dual-colour FISH on paraffin sections
Authors: Bink K, Walch A, Feuchtinger A, Eisenmann H, Hutzler P, Hofler H, Werner M.
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