Thermosensation is crucial for the survival of any organism. In animals, changes in brain temperature are detected via sensory neurons, their cell bodies are located in the trigeminal ganglia. Transient receptor potential (TRP) ion channels are the largest temperature sensing family. In mammals, 11 thermoTRPs are known, as in poultry, there are only three. This research further elucidates TRP mRNA expression in the brain of broiler embryo's. Three incubation treatments were conducted on 400 eggs each: the control (C) at 37.6 °C; T1 deviating from C by providing a + 1 °C heat stimuli during embryonic day (ED) 15–20 for 8 h a day; and T2, imposing a + 2 °C heat stimuli. After each heat stimuli, 12 eggs per treatment were taken for blood sampling from the chorioallantoic membrane and brain harvesting. Incubation parameters such has residual yolk (free embryonic) weight, chick quality and hatch percentage were collected. After primer optimization, 22 target genes (13 TRPs and 9 non-TRPs) were measured on mRNA of the brain using a nanofluidic biochip (Fluidigm Corporation). Four target genes (ANO2, TRPV1, SCN5A, TRAAK) have a significant treatment effect - independent of ED. Another four (TRPM8, TRPA1, TRPM2, TRPC3) have a significant treatment effect visible on one or more ED. Heat sensitive channels were increased in T2 and to a lesser degree in T1, which could be part of an acclimatisation process resulting in later life heat tolerance by increased heat sensitivity. T2, however, resulted in a lower hatch weight, quality and hatchability. No hormonal differences were detected.