Detection of cellular responses to toxicants by dielectrophoresis.
The dielectrophoretic (DEP) crossover method has been applied to the detection of cell responses to toxicants. Time and dose responses of the human cultured leukemia (HL-60) line were measured for paraquat, styrene oxide (SO), N-nitroso-N-methylurea (NMU) and puromycin. These toxicants were chosen because of their different predominant mechanisms of action, namely membrane free radical attack, simultaneous membrane and nucleic acid attack, nucleic acid alkylation, and protein synthesis inhibition, respectively. For all treatments, the specific membrane capacitance (C(mem)) of the cells decreased while the specific membrane conductance (G(mem)) increased in dose- and time-dependent manners. The DEP responses correlated sensitively with alterations in cell surface morphology, especially folds, microvilli, and blebs, observed by scanning electron microscopy. The DEP method was more sensitive to agents that had a direct action on the membrane than to agents for which membrane alterations were secondary. The responses to paraquat and SO, which directly damaged the cell membrane, could be detected 15 min after exposure, while those for puromycin and NMU, which acted on intracellular targets, could be detected after 30 min. The detection times and dose sensitivity results showed that the DEP method is much faster and more sensitive than conventional cell and higher organism viability testing techniques. The feasibility of producing small instruments for toxicity detection and screening based on cellular dielectric responses is discussed.
Ratanachoo
Kanatip
K
Laboratory of Environmental Toxicology, Chulabhorn Research Institute, Vipavadee Rangsit Highway, Lak Si, 10210, Bangkok, Thailand.
Gascoyne
Peter R C
PR
Ruchirawat
Mathuros
M
D002462:Cell Membrane / Q000737:chemistry / Q000187:drug effects* / Q000648:ultrastructure; D002470:Cell Survival / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D025802:Electric Capacitance; D004553:Electric Conductivity; D004586:Electrophoresis / Q000295:instrumentation / Q000379:methods*; D004852:Epoxy Compounds / Q000633:toxicity; D005434:Flow Cytometry; D018922:HL-60 Cells; D006801:Humans; D008770:Methylnitrosourea / Q000633:toxicity; D008832:Microchemistry / Q000379:methods; D008855:Microscopy, Electron, Scanning; D010269:Paraquat / Q000633:toxicity; D011691:Puromycin / Q000633:toxicity; D013499:Surface Properties; D013997:Time Factors; D014116:Toxicology / Q000379:methods*
D004852:Epoxy Compounds; D011691:Puromycin; D008770:Methylnitrosourea; C013690:styrene oxide; D010269:Paraquat