The reliance of parasites on their hosts makes host-parasite interactions ideal models for exploring ecological and evolutionary processes. By providing a consistent supply of parasites, in vivo monocultures offer the opportunity to conduct experiments on a scale that is generally not otherwise possible. Gnathiid isopods are common ectoparasites of marine fishes, and are becoming an increasing focus of research attention due to their experimental amenability and ecological importance as ubiquitous, harmful, blood-feeding "mosquito-like" organisms. They feed on hosts once during each of their three juvenile stages, and after each feeding event they return to the benthos to digest and moult to the next stage. Adults do not feed and remain in the benthos, where they reproduce and give birth. Here, we provide methods of culturing gnathiids, and highlight ways in which gnathiids can be used to examine parasite-host-environment interactions. Captive-raised gnathiid juveniles are increasingly being used in parasitological research; however, the methodology for establishing gnathiid monocultures is still not widely known. Information to obtain in vivo monocultures on teleost fish is detailed for a Great Barrier Reef (Australia) and a Caribbean Sea (US Virgin Islands) gnathiid species, and gnathiid information gained over two decades of successfully maintaining continuous cultures is summarised. Providing a suitable benthic habitat for the predominantly benthic free-living stage of this parasite is paramount. Maintenance comprises provision of adequate benthic shelter, managing parasite populations, and sustaining host health. For the first time, we also measured gnathiids' apparent attack speed (maximum 24.5 cm sec-1; 6.9, 4.9/17.0, median, 25th/75th quantiles) and illustrate how to collect such fast moving ectoparasites in captivity for experiments. In addition to providing details pertaining to culture maintenance, we review research using gnathiid cultures that have enabled detailed scientific understanding of host and parasite biology, behaviour and ecology on coral reefs.