A High Throughput Microplate Feeder Assay for Quantification of Consumption in Drosophila

被引:1
|
作者
Walters, Joshua D. [1 ,2 ]
Hatfield, Jeffrey S. [1 ,2 ]
Baker, Brandon B. [1 ,2 ]
Mackay, Trudy F. C. [1 ,2 ]
Anholt, Robert R. H. [1 ,2 ]
机构
[1] Clemson Univ, Dept Genet & Biochem, Clemson, SC 29631 USA
[2] Clemson Univ, Ctr Human Genet, Clemson, SC 29631 USA
来源
关键词
FOOD;
D O I
10.3791/62771
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Quantifying food intake in Drosophila is used to study the genetic and physiological underpinnings of consumption-associated traits, their environmental factors, and the toxicological and pharmacological effects of numerous substances. Few methods currently implemented are amenable to high throughput measurement. The Microplate Feeder Assay (MFA) was developed for quantifying the consumption of liquid food for individual flies using absorbance. In this assay, flies consume liquid food medium from select wells of a 1536-well microplate. By incorporating a dilute tracer dye into the liquid food medium and loading a known volume into each well, absorbance measurements of the well acquired before and after consumption reflect the resulting change in volume (i.e., volume consumed). To enable high throughput analysis with this method, a 3D-printed coupler was designed that allows flies to be sorted individually into 96-well microplates. This device precisely orients 96- and 1536-well microplates to give each fly access to up to 4 wells for consumption, thus enabling food preference quantification in addition to regular consumption. Furthermore, the device has barrier strips that toggle between open and closed positions to allow for controlled containment and release of a column of samples at a time. This method enables high throughput measurements of consumption of aqueous solutions by many flies simultaneously. It also has the potential to be adapted to other insects and to screen consumption of nutrients, toxins, or pharmaceuticals.
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页数:15
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