Controlling the invasion front of aquatic nuisance species is of high importance to resource managers. We tested the hypothesis that adult sea lamprey (Petromyzon marinus), a destructive invasive species in the Laurentian Great Lakes, would exhibit behavioral avoidance to dual-frequency pulsed direct current generated by vertical electrodes and that the electric field would not injure or kill sea lamprey or non-target fish. Laboratory and in-stream experiments demonstrated that the electric field blocked sea lamprey migration and directed sea lamprey into traps. Rainbow trout (Oncorhynchus mykiss) and white sucker (Catostomus commersoni), species that migrate sympatrically with sea lamprey, avoided the electric field and had minimal injuries when subjected to it. Vertical electrodes are advantageous for fish guidance because (1) the electric field produced varies minimally with depth, (2) the electric field is not grounded, reducing power consumption to where portable and remote deployments powered by solar, wind, hydro, or a small generator are feasible, and (3) vertical electrodes can be quickly deployed without significant stream modification allowing rapid responses to new invasions. Similar dual-frequency pulsed direct current fields produced from vertical electrodes may be advantageous for blocking or trapping other invasive fish or for guiding valued fish around dams.