This material is based on the works supported by Nanyang Assistant Professorship (NAP, M4080740), Agency for Science, Technology and Research (A*STAR) Public Sector Research Funding (PSF, M4070190), A*STAR-JST (The Japan Science and Technology Agency) joint grant (M4070198), and Singapore Ministry of Education (MOE2013-T2-2-049). The authors would like to thank Mr. Roger Tan Kay Chia, Prof. Low Kin Huat, Mr. Poon Kee Chun, Mr. Chew Hock See, Mr. Lam Kim Kheong and Dr. Mao Shixin at School of MAE for their support in setting up and maintaining the research facilities. The authors thank Prof. Michel Maharbiz (U.C. Berkeley) his advice and discussion, Prof. Kris Pister and his group (U.C. Berkeley) for their support in providing the GINA used in this study.

1. Estudio de Animales <ol><li> escarabajos traseros individuales Mecynorrhina torquata (6 cm, 8 g) en envases de plástico por separado con ropa de cama de pellets de madera. </li><li> Alimentar cada escarabajo de una taza de gelatina de azúcar (12 ml) cada 3 días. </li><li> Mantener la temperatura y humedad de la habitación de crianza, a 25 ° C y 60%, respectivamente. </li><li> Prueba de la capacidad de vuelo de cada escarabajo antes de implantar electrodos de alambre fino. <ol><li> tirar suavemente un escarabajo en el aire. Si el escarabajo puede volar durante más de 10 segundos durante 5 ensayos consecutivos, la conclusión de que el escarabajo tiene capacidad de vuelo regulares y emplearla para experimentos posteriores de vuelo. Para recuperar el escarabajo, apagar todas las luces de la habitación para que sea oscuro. Esto hace que el escarabajo de interrumpir el vuelo. Nota: Un escarabajo comienza espontáneamente a volar cuando se liberan en el aire. Es mejor para llevar a cabo los experimentos de vuelo en una habitación grande cerrado tal como el que se muestra en la Figura 1 (16 x 8 x 4 m <sup&gt; 3 con un espacio de captura de movimiento de 12,5 x 8 x 4 m 3), como un escarabajo volador mueve muy rápido (aproximadamente 3-5 m / seg) y atrae a grandes arcos al girar en el aire. </li></ol></li></ol> 2. Electrodo de implantación <ol><li> Anestesiar al escarabajo colocándolo en un recipiente de plástico llena de CO 2 durante 1 min 13,16,20-24. </li><li> Ablandar la cera dental sumergiéndola en agua caliente durante 10 seg. Coloque el escarabajo anestesiado en un bloque de madera e inmovilizarlo con la cera dental ablandada. La cera dental natural se enfría y se solidifica en unos pocos minutos. </li><li> Corte los cables aislados de plata (127 micras de diámetro desnudo, 178 m de diámetro cuando se recubre con perfluoroalkoxy) en longitudes de 25 mm para utilizar como electrodos de alambre fino para la implantación. </li><li> Exponer a 3 mm de plata desnuda por flameado el aislante en ambos extremos de cada cable. </li><li> Diseccionar la superficie superior de la cutícula del escarabajo utilizando una tijera de punta fina para crear un smaventana ll de aproximadamente 4 x 4 mm en la metepisternum (Figura 2c). Nota: una cutícula de color marrón suave se expone entonces, como se muestra en las figuras 2c - e. El músculo 3AX se encuentra debajo de la cutícula suave. </li><li> Pierce dos agujeros en la cutícula marrón expuesto utilizando un PIN de insectos (tamaño 00) con una distancia de 2 mm entre los dos agujeros (Figura 2D). </li><li> Insertar dos electrodos de alambre (incluyendo uno electrodos activos y uno de retorno preparados en la etapa 2.4) con cuidado a través de los agujeros y implantarlos en cada músculo 3AX a una profundidad de 3 mm. </li><li> Asegurar los electrodos implantados y mantenerlos en su lugar para evitar el contacto y cortocircuitos dejando caer cera derretida en los agujeros. Si es necesario, ajustar el flujo de cera de abeja sobre la cutícula tocando la cera de abeja con la punta de un soldador caliente. La cera de abejas se solidifica rápidamente y refuerza la implantación. Nota: Para comprobar si el implante es correcta, los élitros de la remolachaLe puede ser levantado para observar el movimiento del músculo 3AX durante la estimulación eléctrica. </li></ol> 3. Asamblea Mochila inalámbrica Nota: La mochila consistía en un microcontrolador incorporado de radio en una tabla de capas 4 FR-4 (1,6 x 1,6 cm 2). La mochila fue impulsado por un MicroBattery de polímero de litio (3,7 V, 350 mg, 10 mAh). La masa total de la mochila incluyendo la batería fue de 1,2 ± 0,26 g, que es menor que la capacidad de carga útil del escarabajo (30% de 10 g de peso corporal). La mochila se pre-programados para recibir comunicaciones inalámbricas y tenía dos canales de salida. <ol><li> Limpiar la superficie pronoto (quitar la capa de cera sobre la cutícula) con cinta adhesiva de doble cara. A continuación, coloque la mochila en el pronoto del escarabajo con un trozo de cinta de doble cara. </li><li> Conectar los extremos de los electrodos implantados a las salidas de la mochila. </li><li> Envuelva la cinta retrorreflectante alrededor del MicroBattery para producir un marcador for cámaras de captura de movimiento para detectar. </li><li> Coloque la microbatería a la parte superior de la mochila con un trozo de cinta de doble cara para que la cinta retro-reflectante puede ser detectada por las cámaras de captura de movimiento. </li></ol> 4. Sistema de control inalámbrico Nota: En este caso, el sistema de control inalámbrico término incluye un receptor para el mando a distancia, un ordenador portátil para ejecutar el software de control de vuelo a medida, una estación base, la mochila, y el sistema de captura de movimiento. <ol><li> Conectar la estación base y el receptor del mando a distancia para el ordenador portátil a través de puertos USB. </li><li> Encienda el sistema de captura de movimiento y conectarlo a la computadora portátil a través de un puerto Ethernet. </li><li> Realizar la calibración de volumen agitando la varita de calibración (proporcionado por la empresa proveedora del sistema de captura de movimiento) para cubrir la totalidad del espacio de captura de movimiento. <ol><li> Abra el software de captura de movimiento desde el escritorio del ordenador portátil. Haga clic y drag para seleccionar todas las cámaras en el menú &quot;Sistema&quot; del panel &quot;Recursos&quot;. </li><li> Haga clic en el menú &quot;Perspectiva 3D&quot; y seleccione &quot;Cámara&quot; para cambiar a la vista de la cámara. Haga clic en la pestaña &quot;Cámara&quot; en el panel &quot;Herramientas&quot; para mostrar la configuración de calibración. Haga clic en &quot;Inicio&quot; en el menú &quot;Crear máscaras de la cámara&quot; para eliminar el ruido de las cámaras y luego &quot;Stop&quot; después de que el ruido se enmascara en azul. </li><li> Haga clic y seleccione &quot;Marcador de 5 Wand &amp; L-Frame&quot; en el menú &quot;Wand&quot; y el menú &quot;L-Frame&quot; en la pestaña &quot;Cámara&quot;. Ajuste el &quot;Conde Wand&quot; de 2500, haga clic en &quot;Inicio&quot; en el menú &quot;Cámaras Calibrar&quot;, y agitar la varita de calibración a través de todo el espacio de captura de movimiento. El proceso de calibración se detiene cuando el recuento de varita alcanza 2.500. </li><li> Repetir el proceso de calibración de error si la imagen (en la parte inferior de la pestaña &quot;Cámara&quot; del panel &quot;Herramientas&quot;) es superior a 0,3 fo cualquier cámara. Después de la calibración, poner la varita en el suelo en el centro del espacio de captura de movimiento y haga clic en &quot;Inicio&quot; en el menú &quot;Configuración de Origen Volumen&quot; para establecer el origen del espacio de captura de movimiento. </li></ol></li><li> Compruebe la cobertura del sistema de captura de movimiento utilizando una prueba simulada para registrar la trayectoria de movimiento de un marcador ondulado por un usuario en el espacio de captura de movimiento y confirmar si se detecta el marcador y de seguimiento. Si el marcador es frecuentemente pierde durante la detección, repetir la calibración volumen hasta que el maniquí de ensayo tiene éxito. <ol><li> Haga clic en la pestaña de &quot;captura&quot; en el panel &quot;Herramientas&quot; y luego &quot;Inicio&quot; en el menú &quot;Captura&quot; antes de agitar el marcador de la muestra a través de todo el espacio de captura de movimiento para grabar su trayectoria. </li><li> Después de la grabación, haga clic en &quot;Ejecuta la tubería Reconstruir&quot; para reconstruir las posiciones del marcador y comprobar la calidad de la grabación. </li></ol></li><li> Conecte los terminales de la Microbialattery (que se adjunta a la mochila en el paso 3.4) para los pines de alimentación de la mochila. </li><li> Prueba de la comunicación inalámbrica entre el ordenador portátil y la mochila con el software de control de vuelo personalizado. Haga clic en el comando &quot;Inicio&quot; en el software y comprobar el estado de la conexión mostrada. </li></ol> Experimento 5. Vuelo Libre <ol><li> Llevar a cabo el experimento de vuelo libre en una arena de vuelo midiendo 16 x 8 x 4 m 3. </li><li> Introducir los parámetros adecuados en el software de control de vuelo (voltaje, anchura de pulso, frecuencia, y duración de estimulación). Nota: Para la demostración, se fijó la tensión de 3 V, ancho de pulso a 3 mseg, y la duración de estimulación a 1 seg y variada la frecuencia de 60 a 100 Hz. <ol><li> En la pantalla de software, tipo 3 de 3 V en el cuadro de &quot;tensión&quot;, 1.000 por 1.000 mseg en la casilla &quot;Duración de estimulación&quot;, 3 por 3 m en el cuadro de &quot;ancho de pulso&quot;, y una frecuencia deseada en Hz en el &quot; frecuencia &quot;o cuadron la ventana de comandos. </li></ol></li><li> Suelte el escarabajo de la mochila montado en el aire que le permite volar libremente dentro de la arena de vuelo. activar manualmente la estimulación cuando el escarabajo entra en el espacio de captura de movimiento. Pulse el botón correspondiente de comandos (izquierda o derecha) en el control remoto para estimular el músculo deseado en el lado izquierdo o derecho del escarabajo. Nota: Una vez que se pulsa el botón, el software de control de vuelo que se ejecuta en el ordenador portátil genera el comando y lo envía a la mochila. La mochila a continuación, emite el estímulo eléctrico al músculo de interés (en el lado izquierdo o derecho). </li><li> Observar la reacción del escarabajo en tiempo real durante la estimulación y la reconstrucción de los datos utilizando el software de gráficos 3D. <ol><li> Seleccione uno de los ensayos registrados en la lista de datos de la ventana &quot;Display Escarabajo&quot; y haga clic en &quot;Exportar Panda&quot; para copiar los datos de dicho ensayo a la carpeta de análisis y ejecutar el módulo de gráficos 3D. </li><li> Pulse la tecla &quot;N&quot; en lael teclado para combinar la señal de estímulo con la trayectoria grabada. Pulse I para mostrar la trayectoria del escarabajo con los periodos de estimulación resaltados. </li></ol></li></ol>

<ol>
	<li>Kutsch, W., Schwarz, G., Fischer, H., Kautz, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Wireless+Transmission+of+Muscle+Potentials+During+Free+Flight+of+a+Locust.">Wireless Transmission of Muscle Potentials During Free Flight of a Locust.</a> J. Exp. Biol. 185 (1), 367-373 (1993).</li><li>Fischer, H., Kautz, H., Kutsch, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Radiotelemetric+2-Channel+Unit+for+Transmission+of+Muscle+Potentials+During+Free+Flight+of+the+Desert+Locust,+Schistocerca+Gregaria.">A Radiotelemetric 2-Channel Unit for Transmission of Muscle Potentials During Free Flight of the Desert Locust, Schistocerca Gregaria.</a> J. Neurosci. Methods. 64 (1), 39-45 (1996).</li><li>Ando, N., Shimoyama, I., Kanzaki, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Dual-Channel+FM+Transmitter+for+Acquisition+of+Flight+Muscle+Activities+from+the+Freely+Flying+Hawkmoth,+Agrius+Convolvuli.">A Dual-Channel FM Transmitter for Acquisition of Flight Muscle Activities from the Freely Flying Hawkmoth, Agrius Convolvuli.</a> J. Neurosci. Methods. 115 (2), 181-187 (2002).</li><li>Sanchez, C. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Locomotion+control+of+hybrid+cockroach+robots.">Locomotion control of hybrid cockroach robots.</a> J. R. Soc. Interface. 12 (105), (2015).</li><li>Sato, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+cyborg+beetle:+insect+flight+control+through+an+implantable,+tetherless+microsystem.">A cyborg beetle: insect flight control through an implantable, tetherless microsystem.</a> , 164-167 (2008).</li><li>Bozkurt, A., Gilmour, R. F., Lal, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Balloon-Assisted+Flight+of+Radio-Controlled+Insect+Biobots.">Balloon-Assisted Flight of Radio-Controlled Insect Biobots.</a> IEEE Trans. Biomed. Eng. 56 (9), 2304-2307 (2009).</li><li>Sato, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Remote+Radio+Control+of+Insect+Flight.">Remote Radio Control of Insect Flight.</a> Front. Neurosci. 3, (2009).</li><li>Daly, D. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Pulsed+UWB+Receiver+SoC+for+Insect+Motion+Control.">A Pulsed UWB Receiver SoC for Insect Motion Control.</a> IEEE J. Solid-State Circuits. 45 (1), 153-166 (2010).</li><li>Maharbiz, M. M., Sato, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cyborg+Beetles.">Cyborg Beetles.</a> Sci. Am. 303 (6), 94-99 (2010).</li><li>Tsang, W. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Remote+control+of+a+cyborg+moth+using+carbon+nanotube-enhanced+flexible+neuroprosthetic+probe.">Remote control of a cyborg moth using carbon nanotube-enhanced flexible neuroprosthetic probe.</a> , 39-42 (2010).</li><li>Hinterwirth, A. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Wireless+Stimulation+of+Antennal+Muscles+in+Freely+Flying+Hawkmoths+Leads+to+Flight+Path+Changes.">Wireless Stimulation of Antennal Muscles in Freely Flying Hawkmoths Leads to Flight Path Changes.</a> PloS ONE. 7 (12), (2012).</li><li>Whitmire, E., Latif, T., Bozkurt, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Kinect-based+system+for+automated+control+of+terrestrial+insect+biobots.">Kinect-based system for automated control of terrestrial insect biobots.</a> , 1470-1473 (2013).</li><li>Sato, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Deciphering+the+Role+of+a+Coleopteran+Steering+Muscle+via+Free+Flight+Stimulation.">Deciphering the Role of a Coleopteran Steering Muscle via Free Flight Stimulation.</a> Curr. Biol. 25 (6), 798-803 (2015).</li><li>Erickson, J. C., Herrera, M., Bustamante, M., Shingiro, A., Bowen, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effective+Stimulus+Parameters+for+Directed+Locomotion+in+Madagascar+Hissing+Cockroach+Biobot.">Effective Stimulus Parameters for Directed Locomotion in Madagascar Hissing Cockroach Biobot.</a> PLoS ONE. 10 (8), e0134348 (2015).</li><li>Zhaolin, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+preliminary+study+of+motion+control+patterns+for+biorobotic+spiders.">A preliminary study of motion control patterns for biorobotic spiders.</a> , 128-132 (2014).</li><li>Feng, C., Chao, Z., Hao Yu, C., Sato, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Insect-machine+hybrid+robot:+Insect+walking+control+by+sequential+electrical+stimulation+of+leg+muscles.">Insect-machine hybrid robot: Insect walking control by sequential electrical stimulation of leg muscles.</a> , 4576-4582 (2015).</li><li>Cao, F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Biological+Micro+Actuator:+Graded+and+Closed-Loop+Control+of+Insect+Leg+Motion+by+Electrical+Stimulation+of+Muscles.">A Biological Micro Actuator: Graded and Closed-Loop Control of Insect Leg Motion by Electrical Stimulation of Muscles.</a> PLoS ONE. 9 (8), e105389 (2014).</li><li>Zhao, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neuromechanism+Study+of+Insect-Machine+Interface:+Flight+Control+by+Neural+Electrical+Stimulation.">Neuromechanism Study of Insect-Machine Interface: Flight Control by Neural Electrical Stimulation.</a> PLoS ONE. 9 (11), e113012 (2014).</li><li>Tsang, W. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Flexible+Split-Ring+Electrode+for+Insect+Flight+Biasing+Using+Multisite+Neural+Stimulation.">Flexible Split-Ring Electrode for Insect Flight Biasing Using Multisite Neural Stimulation.</a> IEEE Trans. Biomed. Eng. 57 (7), 1757-1764 (2010).</li><li>Barron, A. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anaesthetising+Drosophila+for+behavioural+studies.">Anaesthetising Drosophila for behavioural studies.</a> J. Insect Physiol. 46 (4), 439-442 (2000).</li><li>Cooper, J. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anesthesia,+Analgesia,+and+Euthanasia+of+Invertebrates.">Anesthesia, Analgesia, and Euthanasia of Invertebrates.</a> ILAR Journal. 52 (2), 196-204 (2011).</li><li>Miller, T. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Insect neurophysiological techniques. , (2012).</li><li>Leary, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> AVMA guidelines for the euthanasia of animals. , (2013).</li><li>Heath, B., West, G., Heard, D., Caulkett, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mobile+Inhalant+Anesthesia+Techniques.">Mobile Inhalant Anesthesia Techniques.</a> in Zoo Animal and Wildlife Immobilization and Anesthesia. , 75-80 (2008).</li><li>Mischiati, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Internal+models+direct+dragonfly+interception+steering.">Internal models direct dragonfly interception steering.</a> Nature. 517 (7534), 333-338 (2015).</li><li>Kutsch, W., Berger, S., Kautz, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Turning+Manoeuvres+in+Free-Flying+Locusts:+Two-Channel+Radio-Telemetric+Transmission+of+Muscle+Activity.">Turning Manoeuvres in Free-Flying Locusts: Two-Channel Radio-Telemetric Transmission of Muscle Activity.</a> J. Exp. Zoolog. Part A Comp. Exp. Biol. 299 (2), 139-150 (2003).</li><li>Wang, H., Ando, N., Kanzaki, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Active+Control+of+Free+Flight+Manoeuvres+in+a+Hawkmoth,+Agrius+Convolvuli.">Active Control of Free Flight Manoeuvres in a Hawkmoth, Agrius Convolvuli.</a> J. Exp. Biol. 211 (3), 423-432 (2008).</li><li>Sato, H., Maharbiz, M. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recent+developments+in+the+remote+radio+control+of+insect+flight.">Recent developments in the remote radio control of insect flight.</a> Front. Neurosci. 4, (2010).</li><li>Tien Van, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Flight+behavior+of+the+rhinoceros+beetle+Trypoxylus+dichotomus+during+electrical+nerve+stimulation.">Flight behavior of the rhinoceros beetle Trypoxylus dichotomus during electrical nerve stimulation.</a> Bioinsp. Biomim. 7 (3), 036021 (2012).</li><li>Sane, S. P., Dickinson, M. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+control+of+flight+force+by+a+flapping+wing:+lift+and+drag+production.">The control of flight force by a flapping wing: lift and drag production.</a> J. Exp. Biol. 204 (15), 2607-2626 (2001).</li><li>de Croon, G. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Design,+aerodynamics+and+autonomy+of+the+DelFly.">Design, aerodynamics and autonomy of the DelFly.</a> Bioinsp. Biomim. 7 (2), 025003 (2012).</li><li>Ma, K. Y., Chirarattananon, P., Fuller, S. B., Wood, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Controlled+Flight+of+a+Biologically+Inspired,+Insect-Scale+Robot.">Controlled Flight of a Biologically Inspired, Insect-Scale Robot.</a> Science. 340 (6132), 603-607 (2013).</li></ol>

The authors declare that there are no conflicts of interest.

El proceso de implantación es importante, ya que afecta a la fiabilidad del experimento. Los electrodos deben ser insertados en el músculo a una profundidad de 3 mm o menos, dependiendo del tamaño del escarabajo (evitando el contacto con los músculos cercanos). Si los electrodos tocan los músculos cercanos, acciones motoras y comportamientos indeseables pueden ocurrir debido a la contracción de los músculos cercanos. Los dos electrodos deben estar bien alineados para asegurar que no se produzcan cortocircuitos. C...

An insect-machine hybrid system, often referred to as a cyborg insect or biobot, is the fusion of a living insect platform with a miniature mounted electronic device. The electronic device, which is wirelessly commanded by a remote user, outputs an electrical signal to electrically stimulate neuromuscular sites in the insect via implanted wire electrodes to induce user desired motor actions and behaviors. In the early stages of this research field, researchers were limited to conducting wireless recording of the muscular action of an insect, using simple analog circuits comprised of surface-mounted components1-3. The development of system-on-a-chip technology with radio frequency functionality enabled not only the wireless recording of neuromuscular signals but also the electrical stimulation of the neuromuscular sites in living insects. At present, a built-in radio microcontroller is small enough to be mounted on living insects without causing any obstructions to their locomotion4-13.
The development of the built-in radio microcontroller allows researchers to determine electrical stimulation protocols to induce desired motor actions to control the locomotion of the insect of interest. On the ground, researchers have demonstrated walking control by stimulating the neuromuscular sites of cockroaches4,12,14, spiders15, and beetles16,17. In the air, the initiation and cessation of flight were achieved using different methods such as the stimulation of the optic lobes (the massive neural cluster of a compound eye) in beetles7,9 and brain sub-regions in bees18, whereas turning control has been demonstrated by stimulating the antennae muscles and nervous system of the abdomens in moths11,19 and the flight muscles of beetles7,9,13. In most cases, a built-in radio microcontroller was integrated on a custom-designed printed circuit board to produce a miniature wireless stimulator (backpack), which was mounted on the insect of interest. This allows wireless electrical stimulation to be applied to a freely walking or flying insect. Such a microcontroller-mounted insect is what is referred to as an insect-machine hybrid system. 
This study describes the experimental protocols for building an insect-machine hybrid system, wherein a living beetle is employed as the insect platform, and instructs on how to operate the robot and test its flight control systems. The third axillary sclerite (3Ax) muscle was chosen as the muscle of interest for electrical stimulation and demonstration of left or right turning control13. A pair of thin silver wire electrodes was implanted in both the left and right 3Ax muscles. Moreover, a backpack was mounted on the living beetle. The other ends of the wire electrode were connected to the output pins of the microcontroller. The backpack was small enough for the beetle to carry in flight. Thus, this allows an experimentalist to remotely stimulate the muscle of interest of an insect in free flight and investigate its reactions to the stimulations.

<table><tbody><tr><td>Mecynorrhina torquata beetle</td><td>Kingdom of Beetle Taiwan</td><td></td><td>10 g, 8 cm, pay load capacity is 30% of the body mass&lt;br/&gt; Aproval of importing and using by Agri-Food and Veterinary Authority of Singapore (AVA; HS code: 01069000, product code: ALV002).</td></tr><tr><td>Wireless backpack stimulator</td><td>Custom</td><td></td><td>TI CC2431 micocontroler&lt;br/&gt; The board is custom made based on the GINA board from Prof. Kris Pister&amp;rsquo;s lab. The layout of GINA board can be found at&amp;nbsp;&amp;nbsp;&amp;nbsp; https://openwsn.atlassian.net/wiki/display/OW/GINA</td></tr><tr><td>Wii Remote control</td><td>Nintendo</td><td></td><td>Bluetooth remote control to send the command to the operator laptop</td></tr><tr><td>BeetleCommander v1.8</td><td>Custom. Maharbiz group at UC Berkeley and Sato group at NTU</td><td></td><td>Establish the wireless communication of the backpack and the operator laptop. Configure the stimulus parameters and log the positional data. Visualize the flight data.</td></tr><tr><td>GINA base station</td><td>Kris Pister group at UC Berkeley</td><td></td><td>TI MSP430F2618 and AT86RF231</td></tr><tr><td>Motion capture system</td><td>VICON</td><td>T160</td><td>8 cameras for a flight arena of 12.5 m x 8 m x 4 m</td></tr><tr><td>Motion capture system</td><td>VICON</td><td>T40s</td><td>12 cameras for a flight arena of 12.5 x 8 x 4 m</td></tr><tr><td>Micro battery</td><td>Fullriver</td><td>&amp;nbsp;201013HS10C&amp;nbsp;</td><td>3.7V, 10 mAh</td></tr><tr><td>Retro reflective tape</td><td>Reflexite</td><td>V92-1549-010150</td><td>V92 reflective tape, silver color</td></tr><tr><td>PFA-Insulated Silver Wire&amp;nbsp;</td><td>A-M systems</td><td>786000</td><td>127 &amp;micro;m bare, 177.8 &amp;micro;m coated, 3 mm bare silver flame exposed at tips</td></tr><tr><td>SMT Micro Header&amp;nbsp;</td><td>SAMTEC</td><td>FTSH-110-01-L-DV</td><td>0.3 mm x 6 mm, bend to make a 3 mm long slider to secure the electrode into the PCB header.</td></tr><tr><td>Beeswax</td><td></td><td></td><td>Secure the electrodes</td></tr><tr><td>Dental Wax</td><td>Vertex</td><td></td><td>Immobilize the beetle</td></tr><tr><td>Insect pin</td><td>ROBOZ</td><td>RS-6082-30</td><td>Size&amp;nbsp; 00; 0.3 mm Rod diameter; 0.03 mm tip width; 38 mm Length&amp;nbsp;&lt;br/&gt; Make electrode guiding holes on cuticle</td></tr><tr><td>Tweezers</td><td>DUMONT</td><td>RS-5015</td><td>Pattern #5; .05 mm x .01 mm Tip Size; 110 mm Length&lt;br/&gt; Dissecting and implantation</td></tr><tr><td>Scissors</td><td>ROBOZ</td><td>RS-5620</td><td>Vannas Micro Dissecting Spring Scissors; Straight; 3mm Cutting Edge; 0.1 mm Tip Width; 3&quot; Overall Length&amp;nbsp;&lt;br/&gt; Dissecting and implantation</td></tr><tr><td>Potable soldering iron</td><td>DAIYO</td><td>DS241</td><td>Reflow beeswax</td></tr><tr><td>Hotplate&amp;nbsp;</td><td>CORNING</td><td>PC-400D</td><td>Melting beeswax and dental wax</td></tr><tr><td>Flourescent lamp</td><td>Philips</td><td>TL5 14W</td><td>Light the entire flight arena with 30 panels (60 x 60 cm&lt;sup&gt;2&lt;/sup&gt;). Each panel has 3 lamps.&lt;br/&gt; 14 W, 549 mm x 17 mm&amp;nbsp;</td></tr></tbody></table>

insect-machine hybrid system: remote radio control of a freely flying beetle (mercynorrhina torquata)

El aumento de los dispositivos electrónicos digitales de radio habilitado ha impulsado el uso de pequeñas grabadoras neuromusculares inalámbricas y estimuladores para el estudio de comportamiento de los insectos en vuelo. Esta tecnología permite el desarrollo de un sistema híbrido de insectos-máquina mediante una plataforma de insectos que viven descrito en este protocolo. Además, este protocolo presenta la configuración del sistema y los procedimientos experimentales de vuelo libre para la evaluación de la función de los músculos de vuelo en un insecto sin ataduras. Para la demostración, nos centramos en el tercer músculo axilar sclerite (3AX) para controlar y lograr girando a la izquierda oa la derecha de un escarabajo volador. Un electrodo de alambre de plata fina fue implantado en el músculo 3AX en cada lado del escarabajo. Estos se conectan a las salidas de una mochila inalámbrica (es decir, un estimulador neuromuscular eléctrica) montado en el pronoto del escarabajo. El músculo se estimuló en vuelo libre, alternando el lado de estimulación (izquierda o derecha) o variando la stimulation frecuencia. El escarabajo se volvió hacia el lado ipsilateral cuando el músculo se estimuló y exhibió una respuesta gradual a una frecuencia creciente. El proceso de implantación y calibración del volumen del sistema de cámara de captura de movimiento dimensional 3 deben llevarse a cabo con cuidado para evitar dañar el músculo y perder la pista del marcador, respectivamente. Este método es altamente beneficioso para estudiar el vuelo de insectos, ya que ayuda a revelar las funciones del músculo de vuelo de interés en vuelo libre.

El procedimiento de implantación de electrodos se presenta en la Figura 2 electrodos de hilo de plata fina se implantaron en el músculo 3AX del escarabajo través de pequeños agujeros perforados en la cutícula suave en el músculo (figuras 2d - e).. Esta cutícula suave se encuentra justo por encima del apodema del músculo basalar después de la eliminación de la parte anterior de la metepisternum (Figuras 2d - <st...

Watch this Scientific Journal Video about Insect-machine Hybrid System: Remote Radio Control of a Freely Flying Beetle Mercynorrhina torquata at JoVE.com

Insect-machine Hybrid System: Remote Radio Control of a Freely Flying Beetle Mercynorrhina torquata

This protocol describes the process of constructing an insect-machine hybrid system and carrying out wireless electrical stimulation of the flight muscles required to control the turning motion of a flying insect.

Sistema híbrido de insectos-máquina: radio control remoto de un escarabajo que vuela libremente (<em&gt; Torquata Mercynorrhina</em&gt;)

The rise of radio-enabled digital electronic devices has prompted the use of small wireless neuromuscular recorders and stimulators for studying in-flight ...

insect-machine-hybrid-system-remote-radio-control-freely-flying

Research

JoVE Journal

Neuroscience

Insect-machine Hybrid System: Remote Radio Control of a Freely Flying Beetle (Mercynorrhina torquata)

12.1K vistas.  Nanyang Technological University. This protocol describes the process of constructing an insect-machine hybrid system and carrying out wireless electrical stimulation of the flight muscles required to control the turning motion of a flying insect.

Video: Insect-machine Hybrid System: Remote Radio Control of a Freely Flying Beetle Mercynorrhina torquata

Early Metamorphic Insertion Technology for Insect Flight Behavior Monitoring

Early Metamorphosis Insertion Technology (EMIT) is a novel methodology for integrating microfabricated neuromuscular recording and actuation platforms on insects during their metamorphic development. Here, the implants are fused within the structure and function of the neuromuscular system as a result of metamorphic tissue remaking. The implants emerge with the insect where the development of tissue around the electronics during pupal development results in a bioelectrically and biomechanically enhanced tissue interface. This relatively more reliable and stable interface would be beneficial for many researchers exploring the neural basis of the insect locomotion with alleviated traumatic effects caused during adult stage insertions. In this article, we implant our electrodes into the indirect flight muscles of Manduca sexta. Located in the dorsal-thorax, these main flight powering dorsoventral and dorsolongitudinal muscles actuate the wings and supply the mechanical power for up and down strokes. Relative contraction of these two muscle groups has been under investigation to explore how the yaw maneuver is neurophysiologically coordinated. To characterize the flight dynamics, insects are often tethered with wires and their flight is recorded with digital cameras. We also developed a novel way to tether Manduca sexta on a magnetically levitating frame where the insect is connected to a commercially available wireless neural amplifier. This set up can be used to limit the degree of freedom to yawing &#8220;only&#8221; while transmitting the related electromyography signals from dorsoventral and dorsolongitudinal muscle groups.

We present a novel surgical procedure to implant electrodes in Manduca sexta during its early metamorphic stages. This technique allows mechanically stable and electrically reliable coupling with the neuromuscular tissue to study flight neurophysiology dynamics. We also present a novel magnetic levitation platform for tethered studies of insect yaw.

Tecnología de Inserción Metamórfica temprana para insectos de vuelo Supervisión del comportamiento

Early Metamorphosis Insertion Technology (EMIT) is a novel methodology for integrating microfabricated neuromuscular recording and actuation platforms on ...

Investigación

Comportamiento

Rearing and Double-stranded RNA-mediated Gene Knockdown in the Hide Beetle, Dermestes maculatus

Advances in genomics have raised the possibility of probing biodiversity at an unprecedented scale. However, sequence alone will not be informative without tools to study gene function. The development and sharing of detailed protocols for the establishment of new model systems in laboratories, and for tools to carry out functional studies, is thus crucial for leveraging the power of genomics. Coleoptera (beetles) are the largest clade of insects and occupy virtually all types of habitats on the planet. In addition to providing ideal models for fundamental research, studies of beetles can have impacts on pest control as they are often pests of households, agriculture, and food industries. Detailed protocols for rearing and maintenance of D. maculatus laboratory colonies and for carrying out dsRNA-mediated interference in D. maculatus are presented. Both embryonic and parental RNAi procedures-including apparatus set up, preparation, injection, and post-injection recovery-are described. Methods are also presented for analyzing embryonic phenotypes, including viability, patterning defects in hatched larvae, and cuticle preparations for unhatched larvae. These assays, together with in situ hybridization and immunostaining for molecular markers, make D. maculatus an accessible model system for basic and applied research. They further provide useful information for establishing procedures in other emerging insect model systems.

Rearing and Double-stranded RNA-mediated Gene Knockdown in the Hide Beetle, Dermestes maculatus

Here, we present protocols for rearing an intermediate-germ beetle, Dermestes maculatus (D. maculatus) in the lab. We also share protocols for embryonic and parental RNAi and methods for analyzing embryonic phenotypes to study gene function in this species.

La cría y el doble-stranded RNA mediada gen desmontables en el Hide Escarabajo,<em&gt; Dermestes maculatus</em

Advances in genomics have raised the possibility of probing biodiversity at an unprecedented scale. However, sequence alone will not be informative ...

Genética

Operant Learning of Drosophila at the Torque Meter

For experiments at the torque meter, flies are kept on standard fly medium at 25&deg;C and 60% humidity with a 12hr light/12hr dark regime. A standardized breeding regime assures proper larval density and age-matched cohorts. Cold-anesthetized flies are glued with head and thorax to a triangle-shaped hook the day before the experiment. Attached to the torque meter via a clamp, the fly's intended flight maneuvers are measured as the angular momentum around its vertical body axis. The fly is placed in the center of a cylindrical panorama to accomplish stationary flight. An analog to digital converter card feeds the yaw torque signal into a computer which stores the trace for later analysis. The computer also controls a variety of stimuli which can be brought under the fly's control by closing the feedback loop between these stimuli and the yaw torque trace. Punishment is achieved by applying heat from an adjustable infrared laser.

Measuring the yaw torque of tethered Drosophila with the torque meter allows the neuroscientist exquisite control of the stimulus situation of the experimental animal. Together with the unique genetic tools available in the fruit fly, this paradigm is used for a wide variety of neurobiological research.

El aprendizaje operante de Drosophila en el medidor de torque

For experiments at the torque meter, flies are kept on standard fly medium at 25&deg;C and 60% humidity with a 12hr light/12hr dark regime. A standardized ...

Biología

Measuring the Flight Ability of the Ambrosia Beetle, Platypus Quercivorus (Murayama), Using a Low-Cost, Small, and Easily Constructed Flight Mill

The ambrosia beetle, Platypus quercivorus (Murayama), is the vector of a fungal pathogen that causes mass mortality of Fagaceae trees (Japanese oak wilt). Therefore, knowing the dispersal capacity may help inform trapping/tree removal efforts to prevent this disease more effectively. In this study, we measured the flight velocity and duration and estimated the flight distance of the beetle using a newly developed flight mill. The flight mill is low cost, small, and constructed using commonly available items. Both the flight mill arm and its vertical axis comprise a thin needle. A beetle specimen is glued to one tip of the arm using instant glue. The other tip is thick due to being covered with plastic, thus it facilitates the detection of rotations of the arm. The revolution of the arm is detected by a photo sensor mounted on an infrared LED, and is indicated by a change in the output voltage when the arm passed above the LED. The photo sensor is connected to a personal computer and the output voltage data are stored at a sampling rate of 1 kHz. By conducting experiments using this flight mill, we found that P. quercivorus can fly at least 27 km. Because our flight mill comprises cheap and small ordinary items, many flight mills can be prepared and used simultaneously in a small laboratory space. This enables experimenters to obtain a sufficient amount of data within a short period.

Measuring the Flight Ability of the Ambrosia Beetle, Platypus Quercivorus Murayama, Using a Low-Cost, Small, and Easily Constructed Flight Mill

We developed a low cost and small flight mill, constructed with commonly available items and easily used in experimentation. Using this apparatus, we measured the flight ability of an ambrosia beetle, Platypus quercivorus.

Medición de la capacidad de vuelo del escarabajo de ambrosía, Platypus Quercivorus (Murayama), con un bajo costo, pequeño y vuelo fácilmente construido molino

The ambrosia beetle, Platypus quercivorus (Murayama), is the vector of a fungal pathogen that causes mass mortality of Fagaceae trees (Japanese oak wilt). ...

Medio ambiente

Using Flight Mills to Measure Flight Propensity and Performance of Western Corn Rootworm, Diabrotica virgifera virgifera (LeConte)

The western corn rootworm, Diabrotica virgifera virgifera (LeConte) (Coleoptera: Chrysomelidae), is an economically important pest of corn in the northern United States. Some populations have developed resistance to management strategies including transgenic corn that produces insecticidal toxins derived from the bacterium Bacillus thuringiensis (Bt). Knowledge of western corn rootworm dispersal is of critical importance for models of resistance evolution, spread, and mitigation. Flight behavior of an insect, especially over a long distance, is inherently difficult to observe and characterize. Flight mills provide a means to directly test developmental and physiological impacts and consequences of flight in the laboratory that cannot be obtained in field studies. In this study, flight mills were used to measure the timing of flight activity, total number of flights, and the distance, duration, and speed of flights taken by female rootworms during a 22-h test period. Sixteen flight mills were housed in an environmental chamber with programmable lighting, temperature, and humidity control. The flight mill described is of a typical design, where a flight arm is free to rotate about a central pivot. Rotation is caused by flight of an insect tethered to one end of the flight arm, and each rotation is recorded by a sensor with a time-stamp. Raw data are compiled by software, which are subsequently processed to provide summary statistics for flight parameters of interest. The most difficult task for any flight mill study is attachment of the tether to the insect with an adhesive, and the method used must be tailored to each species. The attachment must be strong enough to hold the insect in a rigid orientation and to prevent detachment during movement, while not interfering with natural wing motion during flight. The attachment process requires dexterity, finesse, and speed, making video footage of the process for rootworms of value.

Using Flight Mills to Measure Flight Propensity and Performance of Western Corn Rootworm, Diabrotica virgifera virgifera LeConte

Flight mills are important tools for comparing how age, sex, mating status, temperature, or various other factors may influence an insect&#8217;s flight behavior. Here we describe protocols to tether and measure the flight propensity and performance of western corn rootworm under different treatments.

Uso de molinos de vuelo para medir la propensión de vuelo y el rendimiento de la raíz de maíz occidental, Diabrotica virgifera virgifera (LeConte)

The western corn rootworm, Diabrotica virgifera virgifera (LeConte) (Coleoptera: Chrysomelidae), is an economically important pest of corn in the northern ...

Extracellular Wire Tetrode Recording in Brain of Freely Walking Insects

Increasing interest in the role of brain activity in insect motor control requires that we be able to monitor neural activity while insects perform natural behavior. We previously developed a technique for implanting tetrode wires into the central complex of cockroach brains that allowed us to record activity from multiple neurons simultaneously while a tethered cockroach turned or altered walking speed. While a major advance, tethered preparations provide access to limited behaviors and often lack feedback processes that occur in freely moving animals. We now present a modified version of that technique that allows us to record from the central complex of freely moving cockroaches as they walk in an arena and deal with barriers by turning, climbing or tunneling. Coupled with high speed video and cluster cutting, we can now relate brain activity to various parameters of the movement of freely behaving insects.

We previously developed a technique for implanting tetrode wires into the central complex of cockroach brains that allows us to monitor activity in individual units of tethered cockroaches. Here we present a modified version of that technique that allows us to also record brain activity in freely moving insects.

Extracelular alambre Tetrodo grabación en el cerebro de Libremente caminar Insectos

Increasing interest in the role of brain activity in insect motor control requires that we be able to monitor neural activity while insects perform ...

Neurociencias

Building an Enhanced Flight Mill for the Study of Tethered Insect Flight

Makerspaces have a high potential of enabling researchers to develop new techniques and to work with novel species in ecological research. This protocol demonstrates how to take advantage of the technology found in makerspaces in order to build a more versatile flight mill for a relatively low cost. Given that this study extracted its&#160;prototype from flight mills built in the last decade, this protocol focuses more on outlining divergences made from the simple, modern flight mill. Previous studies have already shown how advantageous flight mills are to measuring flight parameters such as speed, distance, or periodicity. Such mills have allowed researchers to associate these parameters with morphological, physiological, or genetic factors. In addition to these advantages, this study discusses the benefits of using the technology in makerspaces, like 3D printers and laser cutters, in order to build a more flexible, sturdy, and collapsible flight mill design. Most notably, the 3D printed components of this design allow the user to test insects of various sizes by making the heights of the mill arm and infrared (IR) sensors adjustable. The 3D prints also enable the user to easily disassemble the machine for quick storage or transportation to the field. Moreover, this study makes greater use of magnets and magnetic paint to tether insects with minimal stress. Lastly, this protocol details a versatile analysis of flight data through computer scripts that efficiently separate and analyze differentiable flight trials within a single recording. Although more labor-intensive, applying the tools available in makerspaces and on online 3D modeling programs facilitates multidisciplinary and process-orientated practices and helps researchers avoid costly, premade products with narrowly adjustable dimensions. By taking advantage of the flexibility and reproducibility of technology in makerspaces, this protocol promotes creative flight mill design and inspires open science.

This protocol uses three-dimensional (3D) printers and laser cutters found in makerspaces in order to create a more flexible flight mill design. By using this technology, researchers can reduce costs, enhance design flexibility, and generate reproducible work when constructing their flight mills for tethered insect flight studies.

Construyendo un molino de vuelo mejorado para el estudio del vuelo de insectos atados

Makerspaces have a high potential of enabling researchers to develop new techniques and to work with novel species in ecological research. This protocol ...

Ingeniería

Using Insect Electroantennogram Sensors on Autonomous Robots for Olfactory Searches

Robots designed to track chemical leaks in hazardous industrial facilities1 or explosive traces in landmine fields2 face the same problem as insects foraging for food or searching for mates3: the olfactory search is constrained by the physics of turbulent transport4. The concentration landscape of wind borne odors is discontinuous and consists of sporadically located patches. A pre-requisite to olfactory search is that intermittent odor patches are detected. Because of its high speed and sensitivity5-6, the olfactory organ of insects provides a unique opportunity for detection. Insect antennae have been used in the past to detect not only sex pheromones7 but also chemicals that are relevant to humans, e.g., volatile compounds emanating from cancer cells8 or toxic and illicit substances9-11. We describe here a protocol for using insect antennae on autonomous robots and present a proof of concept for tracking odor plumes to their source. The global response of olfactory neurons is recorded in situ in the form of electroantennograms (EAGs). Our experimental design, based on a whole insect preparation, allows stable recordings within a working day. In comparison, EAGs on excised antennae have a lifetime of 2 hr. A custom hardware/software interface was developed between the EAG electrodes and a robot. The measurement system resolves individual odor patches up to 10 Hz, which exceeds the time scale of artificial chemical sensors12. The efficiency of EAG sensors for olfactory searches is further demonstrated in driving the robot toward a source of pheromone. By using identical olfactory stimuli and sensors as in real animals, our robotic platform provides a direct means for testing biological hypotheses about olfactory coding and search strategies13. It may also prove beneficial for detecting other odorants of interests by combining EAGs from different insect species in a bioelectronic nose configuration14 or using nanostructured gas sensors that mimic insect antennae15.

We describe a protocol for using insect antennae in the form of electroantennograms (EAGs) on autonomous robots. Our experimental design allows stable recordings within a day and resolves individual odor patches up to 10 Hz. The efficiency of EAG sensors for olfactory searches is demonstrated in driving a robot toward an odor source.

El uso de insectos electroantenograma Sensores de Robots Autónoma de Búsquedas olfativas

Robots designed to track chemical leaks in hazardous industrial facilities1 or explosive traces in landmine fields2 face the same problem as insects ...

Radio Frequency Identification and Motion-sensitive Video Efficiently Automate Recording of Unrewarded Choice Behavior by Bumblebees

We present two methods for observing bumblebee choice behavior in an enclosed testing space. The first method consists of Radio Frequency Identification (RFID) readers built into artificial flowers that display various visual cues, and RFID tags (i.e., passive transponders) glued to the thorax of bumblebee workers. The novelty in our implementation is that RFID readers are built directly into artificial flowers that are capable of displaying several distinct visual properties such as color, pattern type, spatial frequency (i.e., &#8220;busyness&#8221; of the pattern), and symmetry (spatial frequency and symmetry were not manipulated in this experiment). Additionally, these visual displays in conjunction with the automated systems are capable of recording unrewarded and untrained choice behavior. The second method consists of recording choice behavior at artificial flowers using motion-sensitive high-definition camcorders. Bumblebees have number tags glued to their thoraces for unique identification. The advantage in this implementation over RFID is that in addition to observing landing behavior, alternate measures of preference such as hovering and antennation may also be observed. Both automation methods increase experimental control, and internal validity by allowing larger scale studies that take into account individual differences. External validity is also improved because bees can freely enter and exit the testing environment without constraints such as the availability of a research assistant on-site. Compared to human observation in real time, the automated methods are more cost-effective and possibly less error-prone.

This video describes Radio-Frequency Identification (RFID) and motion-sensitive video recording methods to monitor choice behavior by bumblebees.

La identificación por radiofrecuencia y video de movimiento sensible de manera eficiente Automatice Grabación de elección no recompensado Comportamiento por abejorros

We present two methods for observing bumblebee choice behavior in an enclosed testing space. The first method consists of Radio Frequency Identification ...

A Simple Flight Mill for the Study of Tethered Flight in Insects

Flight in insects can be long-range migratory flights, intermediate-range dispersal flights, or short-range host-seeking flights. Previous studies have shown that flight mills are valuable tools for the experimental study of insect flight behavior, allowing researchers to examine how factors such as age, host plants, or population source can influence an insects' propensity to disperse. Flight mills allow researchers to measure components of flight such as speed and distance flown. Lack of detailed information about how to build such a device can make their construction appear to be prohibitively complex. We present a simple and relatively inexpensive flight mill for the study of tethered flight in insects. Experimental insects can be tethered with non-toxic adhesives and revolve around an axis by means of a very low friction magnetic bearing. The mill is designed for the study of flight in controlled conditions as it can be used inside an incubator or environmental chamber. The strongest points are the very simple electronic circuitry, the design that allows sixteen insects to fly simultaneously allowing the collection and analysis of a large number of samples in a short time and the potential to use the device in a very limited workspace. This design is extremely flexible, and we have adjusted the mill to accommodate different species of insects of various sizes.

Flight in insects is influenced by a number of factors and the propensity to disperse is an important variable in understanding insect ecology and biological control strategies. We describe the construction and use of a simple, relatively inexpensive, and flexible flight mill for measuring parameters of tethered flight in insects.

Un simple molino de Vuelo para el Estudio de Atado en vuelo Insectos

Flight in insects can be long-range migratory flights, intermediate-range dispersal flights, or short-range host-seeking flights. Previous studies have ...

Sistema híbrido de insectos-máquina: radio control remoto de un escarabajo que vuela libremente (

Resumen

Explorar más videos

Capítulos en este video