• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of biophysjLink to Publisher's site
Biophys J. May 1984; 45(5): 939–946.
PMCID: PMC1434982

Flow cytometric measurement of fluorescence resonance energy transfer on cell surfaces. Quantitative evaluation of the transfer efficiency on a cell-by-cell basis.

Abstract

A method has been developed for the determination of the efficiency (E) of the fluorescence resonance energy transfer between moieties on cell surfaces by use of a computer-controlled flow cytometer capable of dual wavelength excitation. The absolute value of E may be calculated on a single-cell basis. The analysis requires the measurement of samples stained with donor and acceptor conjugated ligands alone as well as together. In model experiments HK 22 murine lymphoma cells labeled with fluorescein-conjugated concanavalin A (Con A) and/or rhodamine conjugated Con A were used to determine energy transfer histograms. Using the analytic solution to energy transfer in two dimensions, a high surface density of Con A binding sites was found that suggests that the Con A receptor sites on the cell surface are to a degree preclustered . We call this technique flow cytometric energy transfer ( FCET ).

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (1.4M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Stryer L. Fluorescence energy transfer as a spectroscopic ruler. Annu Rev Biochem. 1978;47:819–846. [PubMed]
  • Hahn LH, Hammes GG. Structural mapping of aspartate transcarbamoylase by fluorescence energy-transfer measurements: determination of the distance between catalytic sites of different subunits. Biochemistry. 1978 Jun 13;17(12):2423–2429. [PubMed]
  • Zukin RS, Hartig PR, Koshland DE., Jr Use of a distant reporter group as evidence for a conformational change in a sensory receptor. Proc Natl Acad Sci U S A. 1977 May;74(5):1932–1936. [PMC free article] [PubMed]
  • Luedtke R, Owen CS, Vanderkooi JM, Karush F. Proximity relationships within the Fc segment of rabbit immunoglobulin G analyzed by resonance energy transfer. Biochemistry. 1981 May 12;20(10):2927–2936. [PubMed]
  • Epe B, Woolley P, Steinhäuser KG, Littlechild J. Distance measurement by energy transfer: the 3' end of 16-S RNA and proteins S4 and S17 of the ribosome of Escherichia coli. Eur J Biochem. 1982 Dec;129(1):211–219. [PubMed]
  • Damjanovich S, Bähr W, Jovin TM. The functional and fluorescence properties of Escherichia coli RNA polymerase reacted with fluorescamine. Eur J Biochem. 1977 Feb;72(3):559–569. [PubMed]
  • Stryer L, Thomas DD, Meares CF. Diffusion-enhanced fluorescence energy transfer. Annu Rev Biophys Bioeng. 1982;11:203–222. [PubMed]
  • Lobb RR, Auld DS. Stopped-flow radiationless energy transfer kinetics: direct observation of enzyme-substrate complexes at steady state. Biochemistry. 1980 Nov 11;19(23):5297–5302. [PubMed]
  • Thomas DD, Stryer L. Transverse location of the retinal chromophore of rhodopsin in rod outer segment disc membranes. J Mol Biol. 1982 Jan 5;154(1):145–157. [PubMed]
  • Gennis RB, Cantor CR. Use of nonspecific dye labeling for singlet energy-transfer measurements in complex systems. A simple model. Biochemistry. 1972 Jun 20;11(13):2509–2517. [PubMed]
  • Gennis LS, Gennis RB, Cantor CR. Singlet energy-transfer studies on associating protein systems. Distance measurements on trypsin, -chymotrypsin, and their protein inhibitors. Biochemistry. 1972 Jun 20;11(13):2517–2524. [PubMed]
  • Struck DK, Hoekstra D, Pagano RE. Use of resonance energy transfer to monitor membrane fusion. Biochemistry. 1981 Jul 7;20(14):4093–4099. [PubMed]
  • Gingold MP, Rigaud JL, Champeil P. Fluorescence energy transfer between ATPase monomers in sarcoplasmic reticulum reconstituted vesicles. Biochimie. 1981 Nov-Dec;63(11-12):923–925. [PubMed]
  • Dewey TG, Hammes GG. Calculation on fluorescence resonance energy transfer on surfaces. Biophys J. 1980 Dec;32(3):1023–1035. [PMC free article] [PubMed]
  • Estep TN, Thompson TE. Energy transfer in lipid bilayers. Biophys J. 1979 May;26(2):195–207. [PMC free article] [PubMed]
  • Wolber PK, Hudson BS. An analytic solution to the Förster energy transfer problem in two dimensions. Biophys J. 1979 Nov;28(2):197–210. [PMC free article] [PubMed]
  • Snyder B, Freire E. Fluorescence energy transfer in two dimensions. A numeric solution for random and nonrandom distributions. Biophys J. 1982 Nov;40(2):137–148. [PMC free article] [PubMed]
  • Fernandez SM, Berlin RD. Cell surface distribution of lectin receptors determined by resonance energy transfer. Nature. 1976 Dec 2;264(5585):411–415. [PubMed]
  • Chan SS, Arndt-Jovin DJ, Jovin TM. Proximity of lectin receptors on the cell surface measured by fluorescence energy transfer in a flow system. J Histochem Cytochem. 1979 Jan;27(1):56–64. [PubMed]
  • Arndt-Jovin DJ, Grimwade BG, Jovin TM. A dual laser flow sorter utilizing a CW pumped dye laser. Cytometry. 1980 Sep;1(2):127–135. [PubMed]
  • Hardman KD, Wood MK, Schiffer M, Edmundson AB, Ainsworth CF. Structure of concanavalin A at 4.25-ångström resolution. Proc Natl Acad Sci U S A. 1971 Jul;68(7):1393–1397. [PMC free article] [PubMed]
  • Quiocho FA, Reeke GN, Jr, Becker JW, Lipscomb WN, Edelman GM. Structure of soncanavalin A at 4 A resolution. Proc Natl Acad Sci U S A. 1971 Aug;68(8):1853–1857. [PMC free article] [PubMed]
  • Damjanovich S, Trón L, Szöllösi J, Zidovetzki R, Vaz WL, Regateiro F, Arndt-Jovin DJ, Jovin TM. Distribution and mobility of murine histocompatibility H-2Kk antigen in the cytoplasmic membrane. Proc Natl Acad Sci U S A. 1983 Oct;80(19):5985–5989. [PMC free article] [PubMed]
  • Zidovetzki R, Yarden Y, Schlessinger J, Jovin TM. Rotational diffusion of epidermal growth factor complexed to cell surface receptors reflects rapid microaggregation and endocytosis of occupied receptors. Proc Natl Acad Sci U S A. 1981 Nov;78(11):6981–6985. [PMC free article] [PubMed]
  • Hamori E, Arndt-Jovin DJ, Grimwade BG, Jovin TM. Selection of viable cells with known DNA content. Cytometry. 1980 Sep;1(2):132–135. [PubMed]

Articles from Biophysical Journal are provided here courtesy of The Biophysical Society

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

  • Compound
    Compound
    PubChem Compound links
  • MedGen
    MedGen
    Related information in MedGen
  • PubMed
    PubMed
    PubMed citations for these articles
  • Substance
    Substance
    PubChem Substance links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...