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DiTO™-1 [equivalent to TOTO®-1] *5 mM DMSO Solution* *CAS 143413-84-7*

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Physical properties
Molecular weight1302.78
SolventWater
Spectral properties
Extinction coefficient (cm -1 M -1)1170001
Excitation (nm)514
Emission (nm)531
Quantum yield0.341
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
UNSPSC41116134

OverviewpdfSDSpdfProtocol


CAS
143413-84-7
Molecular weight
1302.78
Extinction coefficient (cm -1 M -1)
1170001
Excitation (nm)
514
Emission (nm)
531
Quantum yield
0.341
DiTO™-1 is chemically equivalent to TOTO®-1 (TOTO® is the trademark of Invitrogen). DiTO™-1 is a carbocyanine dimer with green fluorescence similar to FITC. It is cell-impermeant and easily distinguished from Cy5 and rhodamines 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. DiTO™-1 gives strong and selective nuclear staining in cultured cells and in paraffin sections. Simultaneous labeling with cell-permeable Nuclear Red™ LCS1 dye and cell-impermeant TOTO®-1 can be used to assess cell viability. DiTO™-1 and Nuclear Red™ both have much greater extinction coefficients than that of DNA-bound propidium iodide.

Platform


Fluorescence microscope

ExcitationFITC filter set
EmissionFITC filter set
Recommended plateBlack wall/clear bottom
Instrument specification(s)FITC filter set

Example protocol


PREPARATION OF WORKING SOLUTION

DiTO™-1 working solution
Make DiTO™-1 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.

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 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 DiTO™-1 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 FITC filter. 

Calculators


Common stock solution preparation

Table 1. Volume of Water needed to reconstitute specific mass of DiTO™-1 [equivalent to TOTO®-1] *5 mM DMSO Solution* *CAS 143413-84-7* 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 mM76.759 µL383.795 µL767.589 µL3.838 mL7.676 mL
5 mM15.352 µL76.759 µL153.518 µL767.589 µL1.535 mL
10 mM7.676 µL38.379 µL76.759 µL383.795 µL767.589 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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Spectrum


Open in Advanced Spectrum Viewer
spectrum

Spectral properties

Extinction coefficient (cm -1 M -1)1170001
Excitation (nm)514
Emission (nm)531
Quantum yield0.341

Images


Citations


View all 4 citations: Citation Explorer
Biofilm formation on the surface of monazite and xenotime during bioleaching
Authors: van Alin, Arya and Corbett, Melissa K and Fathollahzadeh, Homayoun and Tjiam, M Christian and Rickard, William DA and Sun, Xiao and Putnis, Andrew and Eksteen, Jacques and Kaksonen, Anna H and Watkin, Elizabeth
Journal: Microbial Biotechnology (2023)
Francisella novicida and F. philomiragia biofilm features conditionning fitness in spring water and in presence of antibiotics
Authors: Siebert, Claire and Villers, Corinne and Pavlou, Georgios and Touquet, Bastien and Yakandawala, Nandadeva and Tardieux, Isabelle and Renesto, Patricia
Journal: Plos one (2020): e0228591
Francisella novicida and F. philomiragia biofilm features conditionning fitness in spring water and in presence of antibiotics
Authors: Siebert, Claire and Villers, Corinne and Pavlou, Georgios and Touquet, Bastien and Yakandawala, Nandadeva and Tardieux, Isabelle and Renesto, Patricia
Journal: PloS one (2020): e0228591

References


View all 27 references: Citation Explorer
Chromatin staining of Drosophila testes
Authors: Bonaccorsi S, Giansanti MG, Cenci G, Gatti M.
Journal: Cold Spring Harb Protoc (2012)
In vivo cancer imaging by poly(ethylene glycol)-b-poly(varepsilon-caprolactone) micelles containing a near-infrared probe
Authors: Cho H, Indig GL, Weichert J, Shin HC, Kwon GS.
Journal: Nanomedicine (2012): 228
The fluorescent dyes TO-PRO-3 and TOTO-3 iodide allow detection of microbial cells in soil samples without interference from background fluorescence
Authors: Henneberger R, Birch D, Bergquist P, Walter M, Anitori RP.
Journal: Biotechniques (2011): 190
Effect of Mg ions on efficiency of gene electrotransfer and on cell electropermeabilization
Authors: Haberl S, Miklavcic D, Pavlin M.
Journal: Bioelectrochemistry (2010): 265
Visualizing nuclei in skin cryosections: viable options to 4'6-diamidino-2-phenylindol for confocal laser microscopy
Authors: Glaser K, Wilke K, Wepf R, Biel SS.
Journal: Skin Res Technol (2008): 324
Highly chlorinated Escherichia coli cannot be stained by propidium iodide
Authors: Phe MH, Dossot M, Guilloteau H, Block JC.
Journal: Can J Microbiol (2007): 664
DNA labeling in living cells
Authors: Martin RM, Leonhardt H, Cardoso MC.
Journal: Cytometry A (2005): 45
Quinolinium, 1,1'-[1,3-propanediylbis[(dimethyliminio)-3,1-propanediyl]]bis[4-[3-(3-methyl-2(3 H)-benzothiazolylidene)-1-propen-1-yl]-,iodide (1:4)
Authors: Shan L., undefined
Journal: In: Molecular Imaging and Contrast Agent Database (MICAD), Bethesda (MD). (2004)
Chlorination effect on the fluorescence of nucleic acid staining dyes
Authors: Phe MH, Dossot M, Block JC.
Journal: Water Res (2004): 3729
Sensitive and reliable JC-1 and TOTO-3 double staining to assess mitochondrial transmembrane potential and plasma membrane integrity: interest for cell death investigations
Authors: Zuliani T, Duval R, Jayat C, Schnebert S, Andre P, Dumas M, Ratinaud MH.
Journal: Cytometry A (2003): 100