| Overview |
 Printer Friendly Version
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| Ex/Em (nm) | 560/574 |
| MW | 652.43 |
| CAS # | N/A |
| Solvent | DMSO |
| Storage | F/D/L |
| Category |
Cell Biology Labeling Cells |
| Related |
Viability and Proliferation Cell Viability MDR Research Biochemical Assays |
| Spectrum | Advanced Spectrum Viewer |
1. Prepare 2-10 mM DMSO stock solution
For #22014 add 45 uL DMSO into a 50 µg vial to make 2 mM stock solution (1 mg/ml is equivalent to 1.8 mM);
For #22015 add 36 uL DMSO into a 50 µg vial to make 10 mM stock solution (1 mg/ml is equivalent to 1.46 mM);
For #22016 add 153 uL ml DMSO to make 10 mM stock solution (1 mg/ml is equivalent to 1.53 mM);
For #22017 add 215 uL ml DMSO to make 10 mM stock solution (1 mg/ml is equivalent to 2.15 mM);
For #22020 dissolve 4.2 mg in 1 ml DMSO to make 10 mM stock solution (1 mg/ml is equivalent to ~ 2.4 mM);
Note: The stock solution should be used promptly; any remaining solution should be aliquoted and frozen at < -20 oC. Avoid repeated freeze-thaw cycles, and protect from light.
2. Prepare dye working solution
Prepare a 1 to 20 µM dye working solution right before use by diluting the DMSO stock solution from Step 1 with Hanks and 20 mM Hepes buffer (HHBS) or the buffer of your choice, pH 7. Mix them well by vortexing.
3. Analyze cells with a flow cytometer or a fluorescence microscope:
3.1 Treat cells with test compounds for a desired period of time.
3.2 Centrifuge the cells to get 2-10 x105 cells per tube.
3.3 Resuspend cells in 500 µL of the dye working solution (from Step 2).
3.4 Incubate cells with a dye solution at room temperature or 37 °C for 15 to 30 min, protected from light.
3.5 Remove the dye working solution from the cells, wash the cells with HHBS or buffer of your choice. Resuspend cells in 500 µL of pre-warmed HHBS or medium to get 2-10 x 105 cells per tube.
3.6 Monitor the fluorescence change at Ex/Em = 490/520 nm with a flow cytometer (FL1 channel) or a fluorescence microscope.
Note: For bacterial cells staining: Staining is most efficient when stock solution is diluted 1:800 in nutrient broth preconditioned by overnight growth of the test bacteria, but fresh nutrient broth or PBS may also be used. Bacterial suspensions should be diluted with PBS to 105 - 107 organisms per ml. Bacteria may be stained by applying one ml of solution to .45 µm filter (25mm) and vacuum filtering to remove solution, then adding 1ml of dye solution and incubating 5 - 10 minutes at room temperature.
| References & Citations |
 Printer Friendly Version
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- Zhang C, Jin S, Yang K, Xue X, Li Z, Jiang Y, Chen WQ, Dai L, Zou G, Liang XJ. (2014) Cell membrane tracker based on restriction of intramolecular rotation. ACS Appl Mater Interfaces, 6, 8971.
- Rezatofighi SH, Gould S, Vo BN, Mele K, Hughes WE, Hartley R. (2013) A multiple model probability hypothesis density tracker for time-lapse cell microscopy sequences. Inf Process Med Imaging, 23, 110.
- Klein J, Leupold S, Biegler I, Biedendieck R, Munch R, Jahn D. (2012) TLM-Tracker: software for cell segmentation, tracking and lineage analysis in time-lapse microscopy movies. Bioinformatics, 28, 2276.
- Fang CY, Vaijayanthimala V, Cheng CA, Yeh SH, Chang CF, Li CL, Chang HC. (2011) The exocytosis of fluorescent nanodiamond and its use as a long-term cell tracker. Small, 7, 3363.
- Chalfoun J, Cardone A, Dima AA, Allen DP, Halter MW. (2010) Overlap-Based Cell Tracker. J Res Natl Inst Stand Technol, 115, 477.
- Swirski FK, Berger CR, Figueiredo JL, Mempel TR, von Andrian UH, Pittet MJ, Weissleder R. (2007) A near-infrared cell tracker reagent for multiscopic in vivo imaging and quantification of leukocyte immune responses. PLoS One, 2, e1075.
- Ionescu-Zanetti C, Wang LP, Di Carlo D, Hung P, Di Blas A, Hughey R, Lee LP. (2005) Alkaline hemolysis fragility is dependent on cell shape: results from a morphology tracker. Cytometry A, 65, 116.
- Caetano-Pinto P, Janssen MJ, Gijzen L, Verscheijden L, Wilmer MJ, Masereeuw R. (2016) Fluorescence-Based Transport Assays Revisited in a Human Renal Proximal Tubule Cell Line. Mol Pharm, 13, 933.
- Manna A, De Sarkar S, De S, Bauri AK, Chattopadhyay S, Chatterjee M. (2015) The variable chemotherapeutic response of Malabaricone-A in leukemic and solid tumor cell lines depends on the degree of redox imbalance. Phytomedicine, 22, 713.
- Beem E, Segal MS. (2013) Evaluation of stability and sensitivity of cell fluorescent labels when used for cell migration. J Fluoresc, 23, 975.
