The Rockefeller University, New York, New York 10065, USA.
Department of Biological Sciences, Columbia University in the City of New York, New York, New York 10027, USA
Cold Spring Harb Protoc. 2022 Jun 24;2022(6):Pdb.prot107862. doi: 10.1101/pdb.prot107862.
Both male and female mosquitoes consume sugar-rich nectar meals required for metabolic energy, but only females consume protein-rich blood meals, which are required for egg development. The size of each meal consumed has subsequent effects on behavior and reproduction; therefore, precise quantification is an important aspect of mosquito feeding behavior studies. This protocol describes a high-throughput, end-point assay to quantify meal volumes ingested by individual mosquitoes. The addition of a fluorescent dye to the meal allows for meal size quantification. Individual mosquitoes that have been fed this meal are homogenized in 96-well plates, and the fluorescence levels are measured with a plate reader. This protocol can also be adapted to determine if alteration of meal composition affects the ingested meal volume, if mosquito strain or genotype dictates consumption, or if meals are derived from multiple sources.
雄性和雌性蚊子都需要消耗富含糖分的花蜜来获取代谢能量，但只有雌性蚊子会消耗富含蛋白质的血液来获取卵发育所需的营养。每次进食的量会对蚊子的行为和繁殖产生后续影响；因此，精确的定量分析是蚊子进食行为研究的一个重要方面。本方案描述了一种高通量、终点测定法，可定量测定单个蚊子的进食量。在食物中添加荧光染料可以定量食物的大小。将已经喂食这种食物的个体蚊子在 96 孔板中匀浆，然后用平板读数器测量荧光强度。该方案还可以进行调整，以确定食物成分的改变是否会影响摄入的食物量，是否蚊子的品系或基因型决定了摄入量，或者食物是否来自多个来源。
