As compared to MIT, virus isolation in cell culture is fast and results can be given in 24–48 hours. The cell lines most suitable for virus isolation are of neural origin and the most commonly used cell line is the murine neuroblastoma cell line Neuro-2a. Other cell lines which are used but may not be as sensitive as Neuro-2a include chicken embryo-related (CER) and baby hamster kidney (BHK 21) cells. The suspect clinical specimen or the brain homogenate is inoculated onto the cells grown in a shell vial or 96-well plates, incubated for 24 h and stained by direct FAT after acetone fixation. Recently, a new cell line, human embryonic kidney cell line (HEK 293), was evaluated for isolation of both fixed and street viruses and was found to be as sensitive and specific as Neuro-2a cell line [31] (Figure 3). RTCT is a faster and cheaper alternative to MIT; however it can be performed only in laboratories with cell culture facilities as well as a fluorescent microscope.