Background: Increasing reports of high-resistant Triatominae populations concerns scientists and sanitarians as little is known about the factors behind the occurrence of such phenotype and its real impact on vector control strategies. Moreover, the utilization of a large variety of methodologies hinder the comparison of the reported studies. Methods: This work aims to review laboratory bioassays, redefining the assessed biological features (age, generation and insecticide application area) and technical procedures (mortality recording time and the ideal diagnostic dose). Results: Results were not influenced by the insecticide application area in nymphs or by their generation. Three days-old specimen’s revealed lower susceptibility to the tested insecticide. We determined that it is more appropriate to record mortality 72 h after treatment with insecticide, as well as using a diagnostic dose of 1xDL99. Conclusion: This work suggests more adequate methodological parameters for assessing insecticide resistance in triatomines, which also allows the comparison of results obtained by different research groups. For laboratory bioassays, we recommend: 1) the use of first instar nymphs from first or second generation; 2) 3 day-old specimens; 2) application of insecticide in the dorsal or ventral abdomen area; 3) mortality recording 72 h after treatment with pyrethroids and 4) a diagnostic dose of 1x LD99.