Detection and Purification of Rare Responders by Fixed-Time Flow Cytometry

Chapter
Part of the Springer Lab Manual book series (SLM)

Abstract

Flow cytometrical analysis of cells stained with the calcium-sensitive dye indo-1 (Haugland, 1992) is now widely used for the measurement of cytosolic calcium [Ca2+]i of cells in response to external signals (Tárnok, 1996; 1997; Ulrich et al., 1996). These changes occur typically after stimulation of membrane bound receptors leading to an opening of intracellular calcium stores (Jin et al., 1994) and/or to a change of the activity of calcium channels located in the plasma membrane (Ulrich et al., 1996). These changes in [Ca2+]i are often transient, lasting in the second to minute time range.

Keywords

Surfactant DMSO Serotonin Arginine Pyruvate 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Beisker W (1994) A new combined integral-light and slit-scan data analysis system (DAS) for flow cytometry. Comput Methods Programs Biomed 42: 15 - 26PubMedCrossRefGoogle Scholar
  2. Berridge MJ(1993) Inositol triphosphate and calcium signaling. Nature 361:315-325Google Scholar
  3. Bottenstein JE, Sato G (1979) Growth of a rat neuroblastoma cell line in serum-free sup- plemented medium. Proc Natl Acad Sci USA 76: 514 - 517PubMedCrossRefGoogle Scholar
  4. Dangl JL, Parks DR, Oi VT, Herzenberg, LA (1982) Rapid isolation of immunoglobulin isotype switch variants using fluorescence activated cell sorting. Cytometry 2: 395 - 401PubMedCrossRefGoogle Scholar
  5. Haugland R P (1992) Handbook of fluorescent probes and research chemicals. Molecular Probes Inc., Eugene, OR, USA.Google Scholar
  6. Heumann R, Öcalan M, Kachel V, Hamprecht B (1979) Clonal hybrid cell lines expressing cholinergic and adrenergic properties. Proc Natl Acad Sci USA 76: 4674 - 4677PubMedCrossRefGoogle Scholar
  7. Dunne JF (1991) Time window analysis and sorting. Cytometry 12: 597 - 601PubMedCrossRefGoogle Scholar
  8. Jin W, Loh NM, Loh HH, Thayer SA (1994) Opioids mobilize calcium from inositol 1,4,5 triphosphate-sensitive stores in NG108-15 cells. J Neurosci 14: 1920 - 1929PubMedGoogle Scholar
  9. Julius D, McDermott A B, Axel R, Jessel TM (1988) Molecular characterization of a functional cDNA encoding serotonin receptor. Science 241: 558 - 564PubMedCrossRefGoogle Scholar
  10. Niemann S, Schaller HC (1996) Head-activator and the neuroectodermal differentiation of P19 mouse embryonal carcinoma cells. Neurosci Lett 207: 49 - 52PubMedCrossRefGoogle Scholar
  11. Omann GM, Coppersmith W, Finney DA, Sklar LA (1985) A convenient on-line device for reagent addition, sample mixing, and temperature control of cell suspensions in flow-cytometry. Cytometry 6: 69 - 73PubMedCrossRefGoogle Scholar
  12. Rabinowitch PS, June CH (1994) Intracellular ionized calcium, membrane potential and pH. In: Ormerod MG (ed) Flow cytometry. A Practical Approach, IRL Press, Oxford, pp. 161 - 187Google Scholar
  13. Richter D, Meyerhof W, Buck F, Morley SD (1991) Molecular biology of receptors of neuropeptide hormones. In: Seifert G (ed.) Current topics in pathology: cell receptors. Springer, Berlin, 1991, pp 117 - 139Google Scholar
  14. Schaller HC, Bodenmüller H (1981) Isolation and amino acid sequence of a morpho-genetic peptide in hydra. Proc Natl Acad Sci USA 78: 7000 - 7004PubMedCrossRefGoogle Scholar
  15. Schieren I, MacDermott (1988) A flow cytometric identification and purification of cells by ligand-induced changes in intracellular calcium. J Neurosci Methods 26: 35 - 44Google Scholar
  16. Szöllösi J, Feuerstein BG, Hyun WC, Das, MK, Marton LJ (1991) Attachment of Al27 human glioblastoma cells affects calcium signalling: a comparison of image cytometry, flow cytometry, and spectrofluorometry. Cytometry 12: 707 - 716.PubMedCrossRefGoogle Scholar
  17. Tárnok A (1996) Improved kinetic analysis of cytosolic free calcium in pressure-sensitive neuronal cells by fixed-time flow cytometry. Cytometry 23, pp. 82 - 89.PubMedCrossRefGoogle Scholar
  18. Tárnok A (1997) Rare event sorting based on changes in intracellular free calcium by fixed time flow-cytometry. Cytometry 27: 65 - 70PubMedCrossRefGoogle Scholar
  19. Ulrich H, Tárnok A, Schaller HC (1996) Head-activator induced mitosis of NH15-CA2 cells requires calcium influx and hyperpolarization. J Physiol (Paris) 90: 85 - 94CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2000

Authors and Affiliations

  1. 1.Pediatric CardiologyUniversity Hospital, Herzzentrum Leipzig GmbHLeipzigGermany
  2. 2.Section of Biochemistry, Cell & Molecular BiologyCornell UniversityIthacaUSA

Personalised recommendations