de Castro Noah, Cooper Robin Lewis
Lafayette Senior High School, Lexington, KY 40503, USA.
Department of Biology, University of Kentucky, Lexington, KY 40506-0225, USA.
Methods Protoc. 2020 Jan 24;3(1):12. doi: 10.3390/mps3010012.
Monitoring movements of larval Drosophila with electrical detection allows one to record the behaviors without the use of lights and cameras. This is a suitable technique when studying the use of light-sensitive proteins in optogenetic studies. Electrical measures are feasible to use in determining when a larva starts to move or continues to move after a light induced activation of channelrhodopsin. We have developed a technique using an electrical measure of the media as an index of larval movement. As a proof of concept, recordings with an infrared camera of the larval movement were simultaneous made with electrical measures. The two techniques parallel each other in their ability to index larval movements. Bright light-emitting diode (LED) lights used in optogenetic experiments tend to saturate the detectors of the camera unless filters are used and different filters maybe necessary depending on the LED spectrum and sensitivity of the camera. Impedance measures are independent of the type of LED or brightness. We also assessed the use of a non-solvent based glue (3M Vetbond) to hold larvae in place while measuring synaptic function of neuromuscular junctions, cardiac function and influence of modulators, or activation of light-sensitive channels.
用电检测监测果蝇幼虫的运动,可使人们在不使用灯光和相机的情况下记录行为。在光遗传学研究中研究光敏感蛋白的用途时,这是一种合适的技术。在确定幼虫在光诱导激活通道视紫红质后何时开始移动或继续移动时,电测量是可行的。我们开发了一种技术,利用对培养基的电测量作为幼虫运动的指标。作为概念验证,用红外相机记录幼虫运动的同时进行电测量。这两种技术在索引幼虫运动的能力方面相互平行。光遗传学实验中使用的明亮发光二极管(LED)灯往往会使相机探测器饱和,除非使用滤光片,并且根据LED光谱和相机灵敏度可能需要不同的滤光片。阻抗测量与LED类型或亮度无关。我们还评估了使用非溶剂型胶水(3M Vetbond)在测量神经肌肉接头的突触功能、心脏功能和调节剂的影响或光敏感通道的激活时固定幼虫的情况。