Insect cell-based impedance biosensors: a novel technique to monitor the toxicity of environmental pollutants
Tóm tắt
Cell biosensors are currently emerging as novel, sensitive techniques to monitor the toxicity of environmental pollutants. Here, we have developed electric cell-substrate impedance sensing (ECIS) for on-line monitoring of the behavior of insect cells. Cells were cultured on a microarray of eight small gold electrodes, deposited on the bottom of tissue culture wells. Upon inoculation, cells showed a tendency to drift downward and attached to the gold surface precoated with the protein Concanavalin A to accelerate the cell attachment. The impedance increased because the cells acted as insulating particles to restrict the current flow. The resulting impedance, a coordination of many biological reactions within the cell, was continuously monitored in real-time to reveal information about cell spreading and micromotion. As the cell behavior was sensitive to external chemicals, the applicability of ECIS for inhibition assays was demonstrated with HgCl2, 2,4,6-trinitrotoluene (TNT), 2-amino 4,6-dinitrotoluene (2-ADNT) and 1,3,5-trinitrobenzene (TNB).
Tài liệu tham khảo
Abercrombie M (1982) In: Bellairs R, Curtis A, Dunn, G (eds) Cell behavior. Cambridge University Press, London, pp 10–48
Albrecht-Buehler G (1977) The phagokinetic tracks of 3T3 cells. Cell 11:395–404
Freshney RI (1994) Culture of animal cells: a manual of basic technique, 3rd edn. Wiley-Liss, New York
Ghosh PM, Keese CR, Giaever I (1994) Morphological response of mammalian cells to pulsed ac fields. Bioelectrochem Bioenergetics 33:121–133
Giaever I, Keese CR (1991) Micromotion of mammalian cells measured electrically. Proc Natl Acad Sci USA, Cell Biol 88:7896–7900
Giaever I, Keese CR (1992) Toxic? Cells can tell. Chemtech Feb:116–125
Giaever I, Keese CR (1993) A morphological biosensor for mammalian cells. Nature 366:591–592
Heaysman JEM, Pergum SM (1982) In: Bellairs R, Curtis A, Dunn G (eds) Cell behavior. Cambridge University Press, London, pp 49–76
Keese CR, Giaever I (1994) A biosensor that monitors cell morphology with electrical fields. IEEE Med Biol, June/July:402–408
Keese CR, Karra N, Dillon B, Goldberg AM, Giaever I (1998) Cell-substratum interactions as a predictor of cytotoxicity. In Vitro Mol Toxicol 11:183–192
Ko KS-C, Lo C-M, Ferrier J, Hannam P, Tamura M, McBride BC, Ellen RP (1998) Cell-substrate impedance analysis of epithelial cell shape and micromotion upon challenge with bacterial proteins that perturb extracellular matrix and cytoskeleton. J Microbiol Methods 34:125–132
Kowolenko M, Keese CR, Lawrence DA, Giaever I (1990) Measurement of macrophage adherence and spreading with weal electric fields. J Immunol Methods 127:71–77
Lo C-M, Keese CR, Giaever I (1993) Monitoring motion of confluent cells in tissue culture. Exp Cell Res 204:102–109
Lo C-M, Keese CR, Giaever I (1995) Impedance analysis of MDCK cells measured by electric cell-substrate impedance sensing. Biophys J 69:2800–2807
Luong JHT, Guo Y (1998) Mixed-mode capillary electrokinetic separation of positional explosive isomers using sodium dodecyl sulfate and negative-β-cyclodextrin derivatives. J Chromatogr A 811:225–232
Luong JHT, Mulchandani A, Guilbault GG (1988) Developments and applications of biosensors. Tr Biotechnol 6: 310–316
Luong JHT, Habibi-Rezaei M, Meghrous J, Xiao CD, Male KB, Kamen A (2001) Monitoring motility, spreading, and mortality of adherent insect cells using an impedance sensor. Anal Chem 73:1844–1848
Mitra P, Keese CR, Giaever I (1991) Electric measurements can be used to monitor the attachment and spreading of cells in tissue culture. Biotechniques 11:504–509
Molnar J, Hoekstra S, Ku CS, Van Alten P (1987) Evidence for the recycling nature of the fibronectin receptor of macrophages. J Cell Physiol 131:374–383
Nobel PB, Levine MD (1986) Computer assisted analysis of cell locomotion and chemotaxis. CRC Press, Boca Raton
Pasternak AS, Miller WM (1996) Measurement of trans-epithelial electrical resistance in perfusion: potential application in in vitro ocular toxicity testing. Biotech Bioeng 50:568–579
Scott T, Eagleson M (1988) Concise encyclopedia biochemistry. Walter de Gruyter. Berlin
Smith TJ, Wang HS, Hogg MG, Henrikson RC, Keese CR, Giaever I (1994) Prostaglandin E2 elicits a morphological change in cultured orbital fibroblasts from patients with Graves opthalmopathy. Proc Natl Acad Sci USA 91:5094–5098
Turner APF, Karube I, Wilson GS (1987) Biosensors: fundamentals and applications. Oxford University Press, New York
Xiao C, Lachance B, Sunahara G, Luong JHT (2002) An in-depth analysis of electric cell-substrate impedance sensing to study the attachment and spreading of mammalian cells. Anal Chem 74:1333–1339