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Long wavelength fluorophores and cell-by-cell correction for autofluorescence significantly improves the accuracy of flow cytometric energy transfer measurements on a dual-laser benchtop flow cytometer

  1. Zsolt Sebestyén1,
  2. Péter Nagy2,
  3. Gábor Horváth1,
  4. György Vámosi2,
  5. Reno Debets3,
  6. Jan W. Gratama3,
  7. Denis R. Alexander4,
  8. János Szöllősi1,*

Article first published online: 27 JUN 2002

DOI: 10.1002/cyto.10121

Issue

Cytometry

Cytometry

Volume 48, Issue 3, pages 124–135, 1 July 2002

Additional Information

How to Cite

Sebestyén, Z., Nagy, P., Horváth, G., Vámosi, G., Debets, R., Gratama, J. W., Alexander, D. R. and Szöllősi, J. (2002), Long wavelength fluorophores and cell-by-cell correction for autofluorescence significantly improves the accuracy of flow cytometric energy transfer measurements on a dual-laser benchtop flow cytometer. Cytometry, 48: 124–135. doi: 10.1002/cyto.10121

Author Information

  1. 1

    Department of Biophysics and Cell Biology, Faculty of Medicine, Medical and Health Science Center, University of Debrecen, Debrecen, Hungary

  2. 2

    Cell Biophysics Workgroup of the Hungarian Academy of Sciences, University of Debrecen, Debrecen, Hungary

  3. 3

    Department of Medical Oncology, Clinical and Tumor Immunology, University Hospital Rotterdam–Daniel den Hoed Cancer Center, Rotterdam, The Netherlands

  4. 4

    Laboratory of Lymphocyte Signalling and Development, Programme of Molecular Immunology, The Babraham Institute, Cambridge, United Kingdom

*Department of Biophysics and Cell Biology, Faculty of Medicine, Medical and Health Science Center, University of Debrecen, Nagyerdei kert 98, H-4012 Debrecen, Hungary

Publication History

  1. Issue published online: 27 JUN 2002
  2. Article first published online: 27 JUN 2002
  3. Manuscript Accepted: 1 MAY 2002
  4. Manuscript Revised: 15 APR 2002
  5. Manuscript Received: 1 FEB 2002

Funded by

  • European Commission. Grant Number: FP 5 QLRT-1999-3126
  • Hungarian–British Joint Academic and Research Programme. Grant Number: 069
  • Hungarian Academy of Sciences. Grant Number: OTKA 303999
  • Hungarian Ministry of Health. Grant Numbers: ETT 07-461, ETT 117/2001
  • Hungarian Ministry of Education. Grant Number: FKFP 0607

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LITERATURE CITED

  • 1
    Shivnan E, Biffen M, Shiroo M, Pratt E, Glennie M, Alexander D. Does co-aggregation of the CD45 and CD3 antigens inhibit T cell antigen receptor complex-mediated activation of phospholipase C and protein kinase C? Eur J Immunol 1992; 22: 1055–1062.
    Direct Link:
  • 2
    Stryer L. Fluorescence energy transfer as a spectroscopic ruler. Annu Rev Biochem 1978; 47: 819–846.
  • 3
    Szöllősi J, Damjanovich S, Mátyus L. Application of fluorescence resonance energy transfer in the clinical laboratory: routine and research. Cytometry 1998; 34: 159–179.
    Direct Link:
  • 4
    Hallden G, Skold CM, Eklund A, Forslid J, Hed J. Quenching of intracellular autofluorescence in alveolar macrophages permits analysis of fluorochrome labelled surface antigens by flow cytofluorometry. J Immunol Methods 1991; 142: 207–214.
  • 5
    Beisker W, Dolbeare F, Gray JW. An improved immunocytochemical procedure for high-sensitivity detection of incorporated bromodeoxyuridine. Cytometry 1987; 8: 235–239.
    Direct Link:
  • 6
    Seveus L, Vaisala M, Hemmila I, Kojola H, Roomans GM, Soini E. Use of fluorescent europium chelates as labels in microscopy allows glutaraldehyde fixation and permanent mounting and leads to reduced autofluorescence and good long-term stability. Microsc Res Tech 1994; 28: 149–154.
    Direct Link:
  • 7
    Corsetti JP, Sotirchos SV, Cox C, Cowles JW, Leary JF, Blumburg N. Correction of cellular autofluorescence in flow cytometry by mathematical modeling of cellular fluorescence. Cytometry 1988; 9: 539–547.
    Direct Link:
  • 8
    Alberti S, Bucci C, Fornaro M, Robotti A, Stella M. Immunofluorescence analysis in flow cytometry: better selection of antibody-labeled cells after fluorescence overcompensation in the red channel. J Histochem Cytochem 1991; 39: 701–706.
  • 9
    Alberti S, Parks DR, Herzenberg LA. A single laser method for subtraction of cell autofluorescence in flow cytometry. Cytometry 1987; 8: 114–119.
    Direct Link:
  • 10
    Roederer M, Murphy RF. Cell-by-cell autofluorescence correction for low signal-to-noise systems: application to epidermal growth factor endocytosis by 3T3 fibroblasts. Cytometry 1986; 7: 558–565.
    Direct Link:
  • 11
    Schreiber S, MacDermott RP, Raedler A, Pinnau R, Bertovich MJ, Nash GS. Increased activation of isolated intestinal lamina propria mononuclear cells in inflammatory bowel disease. Gastroenterology 1991; 101: 1020–1030.
  • 12
    Malley A, Stewart CC, Stewart SJ, Waldbeser L, Bradley LM, Shiigi SM. Flow cytometric analysis of I-J expression on murine bone marrow-derived macrophages. J Leukoc Biol 1988; 43: 557–565.
  • 13
    Steinkamp JA, Stewart CC. Dual-laser, differential fluorescence correction method for reducing cellular background autofluorescence. Cytometry 1986; 7: 566–574.
    Direct Link:
  • 14
    Fuchs HJ, McDowell J, Shellito JE. Use of allophycocyanin allows quantitative description by flow cytometry of alveolar macrophage surface antigens present in low numbers of cells. Am Rev Respir Dis 1988; 138: 1124–1128.
  • 15
    Loken MR, Keij JF, Kelley KA. Comparison of helium-neon and dye lasers for the excitation of allophycocyanin. Cytometry 1987; 8: 96–100.
    Direct Link:
  • 16
    Wouters FS, Bastiaens PI, Wirtz KW, Jovin TM. FRET microscopy demonstrates molecular association of non-specific lipid transfer protein (nsL-TP) with fatty acid oxidation enzymes in peroxisomes. EMBO J 1998; 17: 7179–7189.
  • 17
    Bastiaens PI, Majoul IV, Verveer PJ, Soling HD, Jovin TM. Imaging the intracellular trafficking and state of the AB5 quaternary structure of cholera toxin. EMBO J 1996; 15: 4246–4253.
  • 18
    Ng T, Squire A, Hansra G, Bornancin F, Prevostel C, Hanby A, Harris W, Barnes D, Schmidt S, Mellor H, Bastiaens PI, Parker PJ. Imaging protein kinase Cα activation in cells. Science 1999; 283: 2085–2089.
  • 19
    Kenworthy AK. Imaging protein–protein interactions using fluorescence resonance energy transfer microscopy. Methods Comp Methods Enzymol 2001; 24: 289–296.
  • 20
    Trón L, Szöllősi J, Damjanovich S, Helliwell SH, Arndt-Jovin DJ, Jovin TM. Flow cytometric measurement of fluorescence resonance energy transfer on cell surfaces. Quantitative evaluation of the transfer efficiency on a cell-by-cell basis. Biophys J 1984; 45: 939–946.
  • 21
    Brockhoff G, Heiß P, Schegel J, Hofstaedter F, Knuechel R. Epidermal growth factor receptor, c-erbB2 and c-erbB3 receptor interaction, and related cell cycle kinetics of SK-BR-3 and BT474 breast carcinoma cells. Cytometry 2001; 44: 338–348.
    Direct Link:
  • 22
    Hederer RA, Guntermann C, Miller N, Nagy P, Szöllősi J, Damjanovich S, Hale G, Alexander DR. The CD45 tyrosine phosphatase regulates Campath-1H (CD52)–induced TCR-dependent signal transduction in human T cells. Int Immunol 2000; 12: 505–516.
  • 23
    Willemsen RA, Weijtens ME, Ronteltap C, Eshlar Z, Gratama JW, Chames P, Bolhuis RL. Grafting primary human T lymphocytes with cancer-specific chimeric single chain and two chain TCR. Gene Ther 2000; 7: 1369–1377.
  • 24
    Van de Griend RJ, Van Krimpen BA, Bol SJ, Thompson A, Bolhuis RL. Rapid expansion of human cytotoxic T cell clones: growth promotion by a heat-labile serum component and by various types of feeders cells. J Immunol Methods 1984; 10: 285–298.
  • 25
    Szöllősi J, Horejsi V, Bene L, Angelisova P, Damjanovich S. Supramolecular complexes of MHC class I, MHC class II, CD20, and tetraspan molecules (CD53, CD81, and CD82) at the surface of a B cell line JY. J Immunol 1996; 157: 2939–2946.
  • 26
    Bastiaens PI, Jovin TM. Microspectroscopic imaging tracks the intracellular processing of a signal transduction protein: fluorescent-labeled protein kinase C β I. Proc Natl Acad Sci USA 1996; 93: 8407–8412.
  • 27
    Kosk-Kosicka D, Bzdega T, Wawrzynow A. Fluorescence energy transfer studies of purified erythrocyte Ca2+-ATPase. Ca2+-regulated activation by oligomerization. J Biol Chem 1989; 264: 19495–19499.
  • 28
    Aubin JE. Autofluorescence of viable cultured mammalian cells. J Histochem Cytochem 1979; 27: 36–43.
  • 29
    Szöllősi J, Mátyus L, Trón L, Balázs M, Ember I, Fulwyler MJ, Damjanovich S. Flow cytometric measurements of fluorescence energy transfer using single laser excitation. Cytometry 1987; 8: 120–128.
    Direct Link:
  • 30
    Pfeiffer A, Böttcher A, Orso E, Kapinsky M, Nagy P, Bodnár A, Spreitzer I, Liebisch G, Dronbik W, Klaus G, Horn M, Holmer S, Hartung S, Multhoff G, Schütz G, Schindler H, Ulmer AJ, Heine H, Stelter F, Schütt C, Rothe G, Szöllősi J, Damjanovich S, Schmitz G. Lipopolysaccharide and ceramide docking to CD14 provokes ligand-specific receptor clustering in rafts. Eur J Immunol 2001; 31: 3153–3164.
    Direct Link:
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