Journal of Fluorescence

, Volume 23, Issue 5, pp 975–987 | Cite as

Evaluation of Stability and Sensitivity of Cell Fluorescent Labels When Used for Cell Migration

  • Elaine Beem
  • Mark S. SegalEmail author


The directed migration of mammalian cells is a foundation of development and growth. A variety of processes such as tissue development, wound healing, pathogen recognition/destruction as well as cancer metastasis are the result of regulated or dysregulated cell migration. While the ability to measure a cell’s propensity to migrate has clinical relevance in several settings, no universal protocol has been established to measure cell migration. A variety of techniques are currently used to measure migration including manual counting, flow cytometry or Coulter counting, microfluidic devices, computerized spectroscopic methods, or the use of various tracking dyes interfaced with fluorescent or non-fluorescent plate readers. In order to expedite the measurement of migration, we compared several common cytoplasmic and lipophilic cell tracking dyes to determine the best dye for determining migration of rare population of cells. CellVue® Burgundy was found to be superior over calcein AM, Cell Tracker Green CMFDA (chloromethyl fluorescein diacetate), Vybrant CFDA (carboxy fluorescein diacetate succinimidyl ester) in its retention within cells, superior to CellVue® NIR 815, PKH67, and CM DiI with regard to signal to noise ratio, and superior to PKH26 with regard to instrument versatility.


Calcein AM Cell Tracker Green CMFDA (chloromethyl fluorescein diacetate) Vybrant CFDA (carboxy fluorescein diacetate succinimidyl ester) CellVue® NIR 815 PKH26 PKH67 CM DiI CD34 Migration Jurkat 


Source of Funding

NHLBI R01HL079352 to MS.


  1. 1.
    Aiuti A, Webb IJ, Bleul C, Springer T, Gutierrez-Ramos JC (1997) The chemokine SDF-1 is a chemoattractant for human CD34+ hematopoietic progenitor cells and provides a new mechanism to explain the mobilization of CD34+ progenitors to peripheral blood. J Exp Med 185:111–120PubMedCrossRefGoogle Scholar
  2. 2.
    Peled A, Grabovsky V, Habler L, Sandbank J, Arenzana-Seisdedos F, Petit I, Ben-Hur H, Lapidot T, Alon R (1999) The chemokine SDF-1 stimulates integrin-mediated arrest of CD34(+) cells on vascular endothelium under shear flow. J Clin Invest 104:1199–1211PubMedCrossRefGoogle Scholar
  3. 3.
    Asahara T, Masuda H, Takahashi T, Kalka C, Pastore C, Silver M, Kearne M, Magner M, Isner JM (1999) Bone marrow origin of endothelial progenitor cells responsible for postnatal vasculogenesis in physiological and pathological neovascularization. Circ Res 85:221–228PubMedCrossRefGoogle Scholar
  4. 4.
    Dimmeler S, Zeiher AM (2004) Vascular repair by circulating endothelial progenitor cells: the missing link in atherosclerosis? J Mol Med 82:671–677PubMedCrossRefGoogle Scholar
  5. 5.
    Yamaguchi J, Kusano KF, Masuo O et al (2003) Stromal cell-derived factor-1 effects on ex vivo expanded endothelial progenitor cell recruitment for ischemic neovascularization. Circulation 107:1322–1328PubMedCrossRefGoogle Scholar
  6. 6.
    Segal MS, Shah R, Afzal A et al (2006) Nitric oxide cytoskeletal-induced alterations reverse the endothelial progenitor cell migratory defect associated with diabetes. Diabetes 55:102–109PubMedCrossRefGoogle Scholar
  7. 7.
    Essodaigui M, Broxterman HJ, Garnier-Suillerot A (1998) Kinetic analysis of calcein and calcein-acetoxymethylester efflux mediated by the multidrug resistance protein and P-glycoprotein. Biochemistry 37:2243–2250PubMedCrossRefGoogle Scholar
  8. 8.
    Hauser IA, Koziolek M, Hopfer U, Thévenod F (1998) Therapeutic concentrations of cyclosporine A, but not FK506, increase P-glycoprotein expression in endothelial and renal tubule cells. Kidney Int 54:1139–1149PubMedCrossRefGoogle Scholar
  9. 9.
    Thiebaut F, Tsuruo T, Hamada H, Gottesman MM, Pastan I, Willingham MC (1987) Cellular localization of the multidrug-resistance gene product P-glycoprotein in normal human tissues. Proc Natl Acad Sci USA 84:7735–7738PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  1. 1.Division of Nephrology, Hypertension, & Renal Transplantation, Department of MedicineUniversity of FloridaGainesvilleUSA
  2. 2.North Florida/South Georgia Veterans Health SystemGainesvilleUSA

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