Specific pathogen-free ticks infected with tick-borne viruses are required in vector competence studies and studies on tick-virus-host interactions. Infection of ticks by feeding on infected laboratory animals is the only natural way of infection. However, one of the main rules set in ethical guidelines is reduction of the usage of laboratory animals in experiments. Here we describe an alternative method of tick infection with tick-borne encephalitis virus through parenteral inoculation into the tick hemocoel. Although the method bypasses some of the natural barriers in comparison with virus uptake through the tick blood meal, the efficiency of infection is high with over 90% of ticks containing living virus.