We thank Shigeru Matsuyama for providing purified bombykol. This work was supported by the Japan Society for the Promotion of Science KAKENHI (grant numbers 22700197 and 24650090) and the Human Frontier Science Program (HFSP).

1. Experimental Animal <ol><li> Prepare una caja de plástico para mantener las pupas de machos silkmoths (B. mori) hasta su eclosión. Toallas poner papel en la parte inferior y trozos de cartón alrededor de la pared interior de la caja (Figura 1A). Nota: Las piezas de cartón son necesarias para que las polillas adultas de espera mientras se extiende sus alas durante la eclosión (Figura 1A). </li><li> Silkmoth masculina puesto (Bombyx mor i) pupas en la caja y mantenerlos en una incubadora hasta la eclosión debajo de un 16-hr: 8 h luz: oscuridad ciclo a 25 ° C. NOTA: El macho y la hembra pupas pueden ser discriminados por las marcas del sexo en el abdomen (Figura 1B). </li><li> Recoger las polillas macho adultas después de la eclosión y moverlos a una nueva caja. </li><li> Mantenga las polillas adultas en una incubadora bajo una de 16 horas: 8 horas de luz: oscuridad ciclo y disminuir la temperatura a 15 ° C para reducir su actividad antes del experimento. </li></ol> 2. Tethering un Silkmoth <ol><li> La fabricación de un accesorio para la inmovilización (Figura 2A) Nota: El accesorio consiste en un alambre de cobre con una tira de una lámina delgada de plástico en la punta. Esto asegura el movimiento dorsal-ventral del tórax durante la marcha (Figura 2B). <ol><li> Preparar una tira de una lámina delgada de plástico, 2 × 40 mm (grosor: 0,1 mm), y doblar por la mitad. </li><li> Coloque la tira plegada a la punta de un alambre de cobre con un adhesivo. </li><li> Doblar la punta de la tira plegada donde se une el tórax de un Silkmoth. </li></ol></li><li> Utilice polillas adultas (2-8 días de edad) durante el período de luz para el experimento. Nota: La sensibilidad a la feromona depende en gran medida el reloj circadiano 18. Debido B. mori es una polilla diurna, el experimento debe realizarse durante el período de luz. </li><li> Retire con cuidado todas las escalas en el dorSal tórax (mesonoto) con un trozo de tejido húmedo (o un hisopo de algodón) y exponer la cutícula del mesonoto (Figura 2C). </li><li> Pegar un adhesivo en la tira de plástico en la unión y en la superficie de la mesonoto expuesta con un destornillador pequeño de punta plana y esperar 5 a 10 min hasta que el adhesivo ya no es pegajoso. Nota: El adhesivo no debe tocar la bisagra ala o las tégulas ala anterior (Figura 2C). </li><li> Unir el mesonoto a la unión. </li><li> Mantenga la polilla atado antes de colocarlo dentro de la cabina del robot. Mantener el cabezal en un soporte y poner un pedazo de papel debajo de las piernas para descansar la polilla. </li></ol> 3. Robot controlado por insectos <ol><li> Diseñar el hardware del robot insecto controlado basado en trabajos previos 16,17,19. Nota: El robot insecto controlado consiste en una cinta de correr apoyado por el aire con un sensor óptico del ratón para Capture la locomoción de insectos, tableros a base de microcontroladores AVR a medida para el procesamiento y el control motor, y dos motores de corriente continua sin escobillas (Figuras 3 y 4). El robot puede ejecutar sobre la base de la rotación de la bola con una precisión del 96% o superior, dentro de un tiempo de retardo de 200 ms. También asegura la movilidad de máxima velocidad de avance (24,8 mm / seg) y la velocidad angular (96,3 ° / seg) de la Silkmoth durante feromonas comportamiento de seguimiento 16. El flujo de aire de la máquina para correr (Figura 5A) y el sistema de entrega de olor (Figura 5B) están diseñados para la polilla de a bordo para caminar sin problemas en la bola y para adquirir un olor por dos antenas. El canal de admisión de aire y el flujo de la cinta de correr está separado de los del sistema de suministro de olor para evitar la contaminación de la feromona. </li><li> Diseñar el software para los microcontroladores integrados basados en trabajos previos 16. Nota: El microcontrolador a bordo calcula tél movimientos del robot de la locomoción de insectos medida con un sensor óptico (rotacional, Δ X; traslacional, Δ y; Figura 6). La distancia de desplazamiento (Δ L) y ángulo de giro (Δθ) por unidad de tiempo del robot se calculan sobre la base de la distancia de desplazamiento de cada rueda (izquierda, Δ L L; derecha, Δ L R) como Δ L = (Δ L Δ L R) / 2 L + y Δθ = (Δ L L - Δ L R) / rueda D, donde la rueda D es la distancia entre las dos ruedas (120 mm). Δ L L y Δ L R se describen adicionalmente como Δ L L = Δ L x, L + Δ L Y, L y Δ L R = Δ L Δ L x, R + y, R, donde Δ L x, L &lt;/ sub&gt; y Δ L x, R son las distancias de desplazamiento de las ruedas en los lados izquierdo y derecho controladas por Δ Δ x, y L y L, y Δ L y R son los controlados por Δ y. Idealmente, Δ L x, L y L Δ x, R se describen como Δ L x, L = L x -Δ, R = G Δ x (rueda D / bola D), y Δ L Y, L y L Δ y , R se describen como Δ L y, L = Δ L y, R = G Δ y, donde G es la ganancia de motor y la bola D es el diámetro de la bola (50 mm). En la práctica, la ganancia del motor se ajusta de forma independiente por cada lado (izquierdo o derecho de la rueda) y por cada dirección (rotación hacia adelante o hacia atrás) con el fin de calibrar el movimiento del robot. Las ganancias independientes permiten además laajuste de giro del motor asimétrica para generar un sesgo de giro del robot (véase el paso 6.1). </li><li> Lavar la superficie de una bola blanca de poliestireno expandido (masa: aproximadamente 2 g, diámetro: 50 mm) con agua para eliminar cualquier posible señales olfativas o visuales. Nota: La superficie de una nueva bola debe ser maltratado con papel de lija de grano fino, tales como P400, lo que asegura el agarre de las piernas sobre la pelota. </li><li> Encienda el ventilador que suministra aire a las 9 V a la cinta de correr y flota la bola (Figura 5A). Observe la bola del flotador aproximadamente 2 mm desde el fondo de la taza. </li><li> El uso de un tornillo, conecte el cable de cobre del archivo adjunto con la polilla (ver paso 2) a un fijo en la cabina del robot (ver Figura 3 recuadro). Asegúrese de que la posición de las patas del medio está en el centro de la bola (Figura 7A). </li><li> Ajustar la posición vertical de la unión para permitir la polilla de caminar normalmente en la btodas. Mantener la bola a la misma altura antes y después de colocar la polilla (Figura 7B). Nota: Una posición demasiado baja de los datos adjuntos añade presión sobre la polilla y provoca caminar hacia atrás para resistir la presión (Figura 7C), mientras que una posición demasiado alta provoca inestabilidad al caminar y fallos del sensor debido a cambios en la posición vertical de la pelota (Figura 7D). Para comprobar el comportamiento de marcha normal, un solo estímulo-inflado feromona se utiliza para desencadenar caminar en la polilla (para el estímulo de feromonas, consulte el paso 4). Tenga en cuenta que el estímulo de prueba debe ser mínima debido a la exposición previa a bombykol habitúa silkmoths y disminuye su sensibilidad (Matsuyama y Kanzaki, datos no publicados). </li></ol> 4. Preparación de olor Fuente Nota: Hombre B. mori son sensibles a la componente principal de la feromona sexual femenina misma especie (bombykol: (E, Z) -10,12-hexadecadien-1-ol) <shasta&gt; 20. Cualquier contaminación del equipo experimental con bombykol provoca el comportamiento de seguimiento de olor y afecta a la capacidad de respuesta de la polilla. <ol><li> Caída de 10 l de la solución bombykol disuelto en n-hexano (200 ng / l) en un trozo de papel de filtro (aproximadamente 10 mm x 10 mm). La cantidad de bombykol por pieza de papel de filtro es de 2.000 ng. Nota: Para comprobar el comportamiento de marcha normal de la polilla, preparar un cartucho de feromona de estímulo en este paso. El cartucho es un vidrio pipeta Pasteur con una pieza de papel de filtro que contiene 2.000 ng de bombykol. Empujar una bombilla sopla el aire que contiene bombykol. </li></ol> Experimento 5. Olor Fuente Localización <ol><li> Encender el ventilador de un túnel de viento-aire del tipo de tracción (1.800 × 900 × 300 mm, L x W x H; Figura 8) y ajustar la velocidad del viento a 0,7 m / seg. Asegúrese de que la temperatura sea superior a 20 ° C. </li><li> Ajuste la fuente de olor (el pIECE de papel de filtro que contiene bombykol) aguas arriba del túnel de viento. Nota: El ancho de columna se debe confirmar antes del experimento mediante el uso de TiCl4 17,19. </li><li> Encienda la placa de desarrollo del robot y establecer una conexión en serie a un PC a través de Bluetooth. </li><li> Poner en marcha un programa Java a medida llamada &quot;bioseñales&quot;, que proporciona una interfaz entre el PC y el robot. Nota: La ventana principal incluye botones para enviar comandos al robot, ventanas de texto para mostrar la entrada y la salida de la comunicación en serie, y las cajas pequeñas para configurar los parámetros. Los comandos posteriores se envían haciendo clic en los botones correspondientes en este programa, a excepción de la captura de vídeo. </li><li> Haga clic en el botón &quot;Acerca del dispositivo&quot; para confirmar la conexión mediante el envío de un comando al robot a través del puerto COM especificado y comprobar que un mensaje se devuelve por el robot. </li><li> Haga clic en el &quot;memory botón Borrar &quot;para borrar los datos de locomoción anteriores que quedan en la memoria flash interna. </li><li> Haga clic en el botón &quot;drivemode1&quot; para enviar las ganancias de motor por defecto al robot. Nota: Las manipulaciones de las ganancias de motor y el retardo de tiempo entre la locomoción de insectos y movimiento del robot se aplican después de este paso (consulte los pasos 6.1 y 6.3, figura 9). </li><li> Haga clic en el botón &quot;no conduzca&quot; para enviar un comando para inmovilizar el robot hasta que se inicie el experimento. </li><li> Poner el robot en una posición de partida (600 mm más allá de la fuente de olor) y encienda el interruptor de la tarjeta de control del motor. </li><li> Pulsar el botón de grabación de la videocámara para iniciar la captura de vídeo. </li><li> Haga clic en el botón de &quot;inicio de la grabación&quot; para enviar un comando de arranque para iniciar el robot con un registro simultáneo de la rotación de la bola en la memoria flash a bordo. Observe que el robot comienza a moverse y el seguimiento de la pluma de olor. </li><li> Clickea en el&quot;STOP REC&quot; y &quot;no conduzca&quot; botones para enviar comandos para detener tanto el movimiento del robot y la grabación si el robot se localiza la fuente de olor. </li><li> Pulsar el botón de grabación de la videocámara para detener la captura de vídeo. </li><li> Descargar registran los datos de locomoción de la memoria flash a bordo al ordenador a través de una conexión en serie. Cierra el programa. </li></ol> 6. La manipulación del robot controlado por insectos Nota: El tiempo de cada manipulación se indica en la Figura 9. <ol><li> La manipulación de las ganancias de motor Nota: Esta manipulación altera la velocidad de traslación y de rotación del robot. Ganancias de motor asimétricos generan un sesgo torneado, que se puede utilizar para investigar cómo los insectos compensar el sesgo 17. <ol><li> Definir las ganancias de rotación de avance y giro hacia atrás del motor en cada lado 17 (Figura 6B) editando THarchivo de configuración de correo llamado &quot;param2.txt&quot; con un editor de texto. </li><li> Haga clic en el &quot;param2 set&quot; para leer el archivo de configuración modificado en el programa de software. A continuación, haga clic en el &quot;drivemode2&quot; para enviar las ganancias manipulados para el robot. </li></ol></li><li> La inversión de la salida del motor Nota: Esta manipulación proporciona una condición similar a la inversión de la entrada olfativa bilateral (véase el paso 6.4) y se puede utilizar para investigar la importancia de la olfacción bilateral. Sin embargo, la inversión de la salida del motor también se invierte el movimiento visual auto-inducido de una polilla a bordo. El impacto de la entrada visual autoinducida invertida puede evaluarse mediante una comparación con la entrada olfativa invertida 19. <ol><li> Invertir el control motor bilateral mediante el cruce de los cables de control para cada motor. </li></ol></li><li> La manipulación de la demora de tiempo entre la locomoción de insectos y movimiento del robot. Nota: Esta manipulaciónpermite la investigación del periodo aceptable de tiempo empleado en el procesamiento sensorial-motriz para la robótica olor de seguimiento. El microcontrolador almacena los datos de locomoción en una memoria intermedia y procesarlos después del retardo de tiempo especificado. Tenga en cuenta que el robot tiene un retardo de tiempo interna máxima de 200 mseg; Por lo tanto, se espera que el retardo de tiempo real para ser el retardo de tiempo especificado más 200 mseg 16,17. <ol><li> Introducir un número (0-10) en una pequeña caja de la ventana principal para especificar un tiempo de retardo desde 0-1.000 ms en pasos de 100 ms. </li><li> Haga clic en el botón &quot;set retardo&quot; para aplicar el retardo de tiempo. </li></ol></li><li> La manipulación de la entrada olfativa. Nota: Esta manipulación se puede utilizar para investigar el significado de la entrada olfativa bilateral. La dirección oleada de silkmoths está sesgada en el lado de mayor concentración 22. <ol><li> Cambiar la distancia entre la punta del tubo de succión o invertir sus posiciones para alterar eldiferencia en la concentración de olor adquirida por cada antena. </li></ol></li><li> La manipulación de la información visual Nota: Esta manipulación es investigar el papel de la información visual para el olor de seguimiento. <ol><li> Cubra la cubierta con un papel blanco que ocluye 105 ° y 90 ° del campo visual horizontal y vertical de la polilla de a bordo, respectivamente. </li></ol></li></ol>

The authors have nothing to disclose.

Los puntos más importantes para el éxito en el control del robot por un Silkmoth están dejando que la polilla caminar sin problemas en el balón hinchable y la medición de la rotación de la bola de forma estable. Por lo tanto, la inmovilización del Silkmoth y montarlo en la pelota en la posición adecuada son los pasos críticos en este protocolo. adherencia inadecuada de la polilla de la unión o de posicionamiento inadecuado de la polilla en la bola hará que la presión no natural en él, lo que perturba su com...

Autonomous robots capable of finding an odor source can be important for the safety and security of society. They can be used for the detection of disaster victims, of drugs or explosive materials at an airport, and of hazardous material spills or leaks in the environment. At present, we rely entirely on well-trained animals (e.g., dogs) for these tasks, and robotic odor source localization has been strongly expected to relieve the workload of these animals. Finding an odor source is a challenging task for robots because odorants are distributed intermittently in an atmosphere1; therefore, continuous sampling of the odor concentration gradient is not always possible. Thus, a search strategy using intermittent odor cues is necessary for the achievement of robotic odor source localization2-4.
Odor source localization is essential for organism survival and includes tasks such as finding food, mating partners, and sites for oviposition. To overcome the difficulty in tracking patchy distributed odorants, organisms have evolved various behavioral strategies consisting of two fundamental behaviors: moving upstream during odor reception and cross-stream during cessation of odor reception5,6. These reactive strategies have been well-documented in insects and further combined with other modalities, such as wind direction and vision5-8. The insect behavioral models have also been useful examples for robotics3,9-11, in which behavioral algorithms or neural circuit models are implemented into mobile robots for the evaluation of odor source localization abilities10,12-15. From biomimetic perspectives, this "bottom-up" approach is certainly a fundamental way to develop biomimetic robots. However, the bottom-up approach is not a shortcut to obtaining a useful search strategy, because biological analyses are still ongoing, and the modeling of the sensory-motor systems behind insect behaviors has not been completed. Therefore, it is still unknown how such a biological system actually works as a controller of a robotic platform.
In this article, we demonstrate the protocol of a straightforward "top-down" approach to develop an odor-tracking mobile robot controlled by a biological system16,17. The robot is controlled by a real insect and can be regarded as a prototype of future insect-mimetic robots. In the robot's cockpit, a tethered adult male silkmoth (Bombyx mori) walked on an air-supported ball in response to the female sex pheromone, which was delivered to each antenna through air suction tubes. The ball rotations caused by the walking of the onboard moth were measured by an optical sensor and were translated into the movement of the two-wheeled robot. The advantage of this "hybrid" approach is that experimenters can investigate how the insect sensory-motor system works on the robotic platform where a pilot insect is in a closed loop between the robot and a real odor circumstance. The manipulation of the robotic hardware alters the closed loop; therefore, the insect-controlled robot is a useful platform for both engineers and biologists. For engineering, the robot represents the first steps of applying a biological model to meet the requirements for robotic tasks. For biology, the robot is an experimental platform for studying sensory-motor control under a closed loop.

<table><tbody><tr><td>Male adult silkmoth (&lt;em&gt;Bombyx mori&lt;/em&gt;)</td><td></td><td></td><td>Rear from eggs, or purchase as pupae.</td></tr><tr><td>Incubator</td><td>Panasonic</td><td>MIR-254</td><td>Store pupae or adult silkmoths at a constant temperature, 238 L.</td></tr><tr><td>Plastic box</td><td>Sunplatec</td><td>O-3</td><td>Store pupae or adult silkmoths, 299 &amp;times; 224 &amp;times; 62 mm L &amp;times; W &amp;times; H.</td></tr><tr><td>Copper wire</td><td></td><td></td><td>2-mm diameter for the attachment. Any rigid bar can be used as an alternative for making the attachment to tether a silkmoth.&amp;nbsp;</td></tr><tr><td>Plastic sheet</td><td>Kokuyo</td><td>VF-1420N</td><td>Sold as overhead projector film with thickness of 0.1 mm. Use at the tip of the attachment.</td></tr><tr><td>Forceps</td><td>As one</td><td>5SA</td><td>Remove scales on the thorax.</td></tr><tr><td>Adhesive</td><td>Konishi</td><td>G17</td><td>Bond a silkmoth to the attachment.</td></tr><tr><td>Insect-controlled robot</td><td>Custom</td><td></td><td>Bearing an air-supported treadmill, an optical sensor, custom-built AVR-based microcontroller boards, and two DC brushless motors. It is powered by 8 &amp;times; AA and 3 &amp;times; 006P batteries.</td></tr><tr><td>Microcontroller</td><td>Atmel</td><td>ATMEGA8</td><td>A component of the insect-controlled robot.</td></tr><tr><td>DC blower</td><td>Nidec</td><td>A34342-55</td><td>A component of the insect-controlled robot for floating a ball in an air-supported treadmill.&amp;nbsp;</td></tr><tr><td>DC fan</td><td>Minebea</td><td>1606KL-04W-B50</td><td>A component of the insect-controlled robot for suctioning air containing an odor.</td></tr><tr><td>Optical mouse sensor</td><td>Agilent technologies</td><td>HDNS-2000</td><td>A component of the insect-controlled robot, obtained from an optical mouse (M-GUWSRSV, Elecom, Japan).</td></tr><tr><td>Brushless motor</td><td>Maxon</td><td>EC-45</td><td>A component of the insect-controlled robot for driving a wheel.</td></tr><tr><td>White polystyrene ball</td><td></td><td></td><td>A component of the insect-controlled robot. Diameter 50 mm, mass approximately 2 g.</td></tr><tr><td>Bombykol:&lt;br/&gt;(E,Z)-10,12-hexadecadien-1-ol</td><td>Shin-Etsu chemical</td><td></td><td>Custom synthesis.</td></tr><tr><td>n-hexane</td><td>Wako</td><td>085-00416</td><td>Solvent for bombykol.</td></tr><tr><td>Wind tunnel</td><td>Custom</td><td></td><td>Pulling-air type, sized 1,800 &amp;times; 900 &amp;times; 300 mm L &amp;times; W &amp;times; H.</td></tr><tr><td>BioSignal program</td><td>Custom</td><td></td><td>A program to establish serial communication between the insect-controlled robot and a PC via Bluetooth. Used for sending commands to start/stop the robot or configuring its motor properties.&amp;nbsp;</td></tr><tr><td>Camcorder</td><td>Sony</td><td>HDR-XR520V</td><td>Capture robot movements.</td></tr></tbody></table>

insect-controlled robot: a mobile robot platform to evaluate the odor-tracking capability of an insect

Robotic odor source localization has been a challenging area and one to which biological knowledge has been expected to contribute, as finding odor sources is an essential task for organism survival. Insects are well-studied organisms with regard to odor tracking, and their behavioral strategies have been applied to mobile robots for evaluation. This "bottom-up" approach is a fundamental way to develop biomimetic robots; however, the biological analyses and the modeling of behavioral mechanisms are still ongoing. Therefore, it is still unknown how such a biological system actually works as the controller of a robotic platform. To answer this question, we have developed an insect-controlled robot in which a male adult silkmoth (Bombyx mori) drives a robot car in response to odor stimuli; this can be regarded as a prototype of a future insect-mimetic robot. In the cockpit of the robot, a tethered silkmoth walked on an air-supported ball and an optical sensor measured the ball rotations. These rotations were translated into the movement of the two-wheeled robot. The advantage of this "hybrid" approach is that experimenters can manipulate any parameter of the robot, which enables the evaluation of the odor-tracking capability of insects and provides useful suggestions for robotic odor-tracking. Furthermore, these manipulations are non-invasive ways to alter the sensory-motor relationship of a pilot insect and will be a useful technique for understanding adaptive behaviors.

Presentamos aquí las características básicas del robot insecto controlado requerida para la localización exitosa de una fuente de olor. La comparación entre el robot y silkmoths, la eficacia del sistema de suministro de olor, y la importancia de olfativo bilateral exacto y entradas visuales son examinados. La comparación de los comportamientos de olor de seguimiento entre polillas libremente a pie y el robot de insectos ...

Watch this Scientific Journal Video about Insect-controlled Robot: A Mobile Robot Platform to Evaluate the Odor-tracking Capability of an Insect at JoVE.com

Insect-controlled Robot: A Mobile Robot Platform to Evaluate the Odor-tracking Capability of an Insect

La capacidad para localizar una fuente de olor es necesario para la supervivencia de insectos y se espera que sea aplicable a artificial olor de seguimiento. El robot insecto controlado es impulsado por un Silkmoth real y nos permite evaluar la capacidad de seguimiento de olor de los insectos a través de una plataforma robótica.

Robot controlado por insectos: una plataforma del robot móvil para evaluar la capacidad de seguimiento de Olor de un insecto

Robotic odor source localization has been a challenging area and one to which biological knowledge has been expected to contribute, as finding odor ...

insect-controlled-robot-mobile-robot-platform-to-evaluate-odor

Universidad de Cartagena UDEC

Research

JoVE Journal

Neuroscience

14.6K vistas.  The University of Tokyo.  La capacidad para localizar una fuente de olor es necesario para la supervivencia de insectos y se espera que sea aplicable a artificial olor de seguimiento. El robot insecto controlado es impulsado por un Silkmoth real y nos permite evaluar la capacidad de seguimiento de olor de los insectos a través de una plataforma robótica. 

Video: Insect-controlled Robot: A Mobile Robot Platform to Evaluate the Odor-tracking Capability of an Insect

High-resolution Measurement of Odor-Driven Behavior in Drosophila Larvae

Olfactory responses in Drosophila larvae have been traditionally studied in Petri dishes comprising a single peripheral odor source. In this behavioral paradigm, the experimenter usually assumes that the rapid diffusion of odorant molecules from the source leads to the creation of a stable gradient in the dish. To establish a quantitative correlation between sensory inputs and behavioral responses, it is necessary to achieve a more thorough characterization of the odorant stimulus conditions. In this video article, we describe a new method allowing the construction of odorant gradients with stable and controllable geometries. We briefly illustrate how these gradients can be used to screen for olfactory defects (full and partial anosmia) and to study more subtle features of chemotaxis behavior.

In this video article, we describe a new method allowing the construction of odorant gradients with stable and controllable geometries. We briefly illustrate how these gradients can be used to screen for olfactory defects (full and partial anosmia) and to study more subtle features of chemotaxis behavior.

De alta resolución de medición del comportamiento de olor-Driven en larvas de Drosophila

Olfactory responses in Drosophila larvae have been traditionally studied in Petri dishes comprising a single peripheral odor source. In this behavioral ...

Investigación

Biología

Visually Mediated Odor Tracking During Flight in Drosophila

Flying insects use visual cues to stabilize their heading in a wind stream. Many animals additionally track odors carried in the wind. As such, visual stabilization of upwind tracking directly aids in odor tracking. But do olfactory signals directly influence visual tracking behavior independently from wind cues? Additionally, recent advances in olfactory molecular genetics and neurophysiology have motivated novel quantitative behavioral analyses to assess the behavioral influence of (e.g.) genetically inactivating specific olfactory activation circuits. We modified a magnetic tether system originally devised for vision experiments by equipping the arena with narrow laminar flow odor plumes. Here we focus on experiments that can be performed after a fly is tethered and is able to navigate in the magnetic arena. We show how to acquire video images optimized for measuring body angle, how to judge stable odor tracking, and we illustrate two experiments to examine the influence of visual cues on odor tracking.

Here we describe how to optimize the acquired video image for an olfactory magnetic-tether (OMT) apparatus. We also describe two sample experimental protocols for studying visuo-olfactory fusion.

Seguimiento visual mediada olor durante el vuelo en Drosophila

Flying insects use visual cues to stabilize their heading in a wind stream. Many animals additionally track odors carried in the wind. As such, visual ...

A Wind Tunnel for Odor Mediated Insect Behavioural Assays

Olfaction is the most important sensory mechanism by which many insects interact with their environment and a wind tunnel is an excellent tool to study insect chemical ecology. Insects can locate point sources in a three-dimensional environment through the sensory interaction and sophisticated behavior. The quantification of this behavior is a key element in the development of new tools for pest control and decision support. A wind tunnel with a suitable flight section with laminar air flow, visual cues for in-flight feedback and a variety of options for the application of odors can be used to measure complex behaviour which subsequently may allow the identification of attractive or repellent odors, insect flight characteristics, visual-odor interactions and interactions between attractants and odors lingering as background odors in the environment. A wind tunnel holds the advantage of studying the odor mediated behavioural repertoire of an insect in a laboratory setting. Behavioural measures in a controlled setting provide the link between the insect physiology and field application. A wind tunnel must be a flexible tool and should easily support the changes to setup and hardware to fit different research questions. The major disadvantage to the wind tunnel setup described here, is the clean odor background which necessitates special attention when developing a synthetic volatile blend for field application.

Here, we describe the construction and use of a wind tunnel for odor mediated behavioural assays with insects. The wind tunnel design facilitates the release of odor sources by several methods, with and without visual stimuli. Wind tunnel experiments are important methods to identify behaviorally active volatile chemicals.

Un túnel de viento para olor mediada por insectos ensayos conductuales

Olfaction is the most important sensory mechanism by which many insects interact with their environment and a wind tunnel is an excellent tool to study ...

Comportamiento

Flying Insect Detection and Classification with Inexpensive Sensors

An inexpensive, noninvasive system that could accurately classify flying insects would have important implications for entomological research, and allow for the development of many useful applications in vector and pest control for both medical and agricultural entomology. Given this, the last sixty years have seen many research efforts devoted to this task. To date, however, none of this research has had a lasting impact. In this work, we show that pseudo-acoustic optical sensors can produce superior data; that additional features, both intrinsic and extrinsic to the insect&#8217;s flight behavior, can be exploited to improve insect classification; that a Bayesian classification approach allows to efficiently learn classification models that are very robust to over-fitting, and a general classification framework allows to easily incorporate arbitrary number of features. We demonstrate the findings with large-scale experiments that dwarf all previous works combined, as measured by the number of insects and the number of species considered.

We proposed a system that uses inexpensive, noninvasive pseudo-acoustic optical sensors to automatically and accurately detect, count, and classify flying insects based on their flying sound.

Flying detección de insectos y Clasificación con Sensores Barato

An inexpensive, noninvasive system that could accurately classify flying insects would have important implications for entomological research, and allow ...

Bioingeniería

Vertical T-maze Choice Assay for Arthropod Response to Odorants

Given the economic importance of insects and arachnids as pests of agricultural crops, urban environments or as vectors of plant and human diseases, various technologies are being developed as control tools. A subset of these tools focuses on modifying the behavior of arthropods by attraction or repulsion. Therefore, arthropods are often the focus of behavioral investigations. Various tools have been developed to measure arthropod behavior, including wind tunnels, flight mills, servospheres, and various types of olfactometers. The purpose of these tools is to measure insect or arachnid response to visual or more often olfactory cues. The vertical T-maze oflactometer described here measures choices performed by insects in response to attractants or repellents. It is a high throughput assay device that takes advantage of the positive phototaxis (attraction to light) and negative geotaxis (tendency to walk or fly upward) exhibited by many arthropods. The olfactometer consists of a 30 cm glass tube that is divided in half with a Teflon strip forming a T-maze. Each half serves as an arm of the olfactometer enabling the test subjects to make a choice between two potential odor fields in assays involving attractants. In assays involving repellents, lack of normal response to known attractants can also be measured as a third variable.

A vertical, T-maze olfactometer is described for assaying the behavioral response of arthropods. The olfactometer allows the experimenter to measure choices performed by test subjects when subjected to two potential odor fields. Both attraction to and repulsion from odorants can be measured with this device.

Vertical T-laberinto Choice Ensayo para la respuesta a los olores artrópodos

Given the economic importance of insects and arachnids as pests of agricultural crops, urban environments or as vectors of plant and human diseases, ...

Neurociencias

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 ...

Electroantennography-based Bio-hybrid Odor-detecting Drone using Silkmoth Antennae for Odor Source Localization

Small drones with chemical or biosensor devices that can detect airborne odorant molecules have attracted considerable attention owing to their applicability in environmental and security monitoring and search-and-rescue operations. Small drones with commercial metal-oxide-semiconductor (MOX) gas sensors have been developed for odor source localization; however, their real-time-odor-detection performance has proven inadequate. However, biosensing technologies based on insect olfactory systems exhibit relatively high sensitivity, selectivity, and real-time response with respect to odorant molecules compared to commercial MOX gas sensors. In such devices, excised insect antennae function as portable odorant biosensor elements and have been found to deliver excellent sensing performance. This study presents experimental protocols for odorant-molecule detection in the air using a small autonomous bio-hybrid drone based on a mountable electroantennography (EAG) device incorporating silkmoth antennae.
We developed a mountable EAG device including sensing/processing parts with a Wi-Fi module. The device was equipped with a simple sensor enclosure to enhance the sensor directivity. Thus, odor source localization was conducted using the spiral-surge algorithm, which does not assume an upwind direction. The experimental bio-hybrid odor-detecting drone identified real-time odorant-concentration differences in a pseudo-open environment (outside a wind tunnel) and localized the source. The developed drone and associated system can serve as an efficient odorant molecule-detection tool and a suitable flight platform for developing odor source localization algorithms owing to its high programmability.

This study introduces experimental protocols for a bio-hybrid odor-detecting drone based on silkmoth antennae. The operation of an experimental electroantennogram device with silkmoth antennae is presented, in addition to the structure of a bio-hybrid drone designed for odor source localization using the spiral-surge algorithm.

Drone bio-híbrido de detección de olores basado en electroantennografía utilizando antenas Silkmoth para la localización de fuentes de olor

Small drones with chemical or biosensor devices that can detect airborne odorant molecules have attracted considerable attention owing to their ...

Ingeniería

High-resolution Quantification of Odor-guided Behavior in Drosophila melanogaster Using the Flywalk Paradigm

In their natural environment, insects such as the vinegar fly Drosophila melanogaster are bombarded with a huge amount of chemically distinct odorants. To complicate matters even further, the odors detected by the insect nervous system usually are not single compounds but mixtures whose composition and concentration ratios vary. This leads to an almost infinite amount of different olfactory stimuli which have to be evaluated by the nervous system.
To understand which aspects of an odor stimulus determine its evaluation by the fly, it is therefore desirable to efficiently examine odor-guided behavior towards many odorants and odor mixtures. To directly correlate behavior to neuronal activity, behavior should be quantified in a comparable time frame and under identical stimulus conditions as in neurophysiological experiments. However, many currently used olfactory bioassays in Drosophila neuroethology are rather specialized either towards efficiency or towards resolution.
Flywalk, an automated odor delivery and tracking system, bridges the gap between efficiency and resolution. It allows the determination of exactly when an odor packet stimulated a freely walking fly, and to determine the animal&#180;s dynamic behavioral reaction.

High-resolution Quantification of Odor-guided Behavior in Drosophila melanogaster Using the Flywalk Paradigm

The automated tracking system Flywalk is used for high-resolution quantification of odor-guided behavior in Drosophila melanogaster.

De alta resolución de cuantificación del comportamiento guiado Olor en<em&gt; Drosophila melanogaster</em&gt; Uso de la<em&gt; Flywalk</em&gt; Paradigma

In their natural environment, insects such as the vinegar fly Drosophila melanogaster are bombarded with a huge amount of chemically distinct odorants. To ...

Olfactory Behaviors Assayed by Computer Tracking Of Drosophila in a Four-quadrant Olfactometer

A key challenge in neurobiology is to understand how neural circuits function to guide appropriate animal behaviors. Drosophila melanogaster is an excellent model system for such investigations due to its complex behaviors, powerful genetic techniques, and compact nervous system. Laboratory behavioral assays have long been used with Drosophila to simulate properties of the natural environment and study the neural mechanisms underlying the corresponding behaviors (e.g. phototaxis, chemotaxis, sensory learning and memory)1-3. With the recent availability of large collections of transgenic Drosophila lines that label specific neural subsets, behavioral assays have taken on a prominent role to link neurons with behaviors4-11. Versatile and reproducible paradigms, together with the underlying computational routines for data analysis, are indispensable for rapid tests of candidate fly lines with various genotypes. Particularly useful are setups that are flexible in the number of animals tested, duration of experiments and nature of presented stimuli. The assay of choice should also generate reproducible data that is easy to acquire and analyze. Here, we present a detailed description of a system and protocol for assaying behavioral responses of Drosophila flies in a large four-field arena. The setup is used here to assay responses of flies to a single olfactory stimulus; however, the same setup may be modified to test multiple olfactory, visual or optogenetic stimuli, or a combination of these. The olfactometer setup records the activity of fly populations responding to odors, and computational analytical methods are applied to quantify fly behaviors. The collected data are analyzed to get a quick read-out of an experimental run, which is essential for efficient data collection and the optimization of experimental conditions.

Olfactory Behaviors Assayed by Computer Tracking Of Drosophila in a Four-quadrant Olfactometer

We describe here a behavioral setup and data analysis method for assaying olfactory responses of up to 100 vinegar flies (Drosophila melanogaster). This system may be used with single or multiple olfactory stimuli, and adaptable for optogenetic activation or silencing of neuronal subsets.

Comportamientos olfativas ensayó mediante seguimiento informático de<em&gt; Drosophila</em&gt; En un olfatómetro de cuatro cuadrantes

A key challenge in neurobiology is to understand how neural circuits function to guide appropriate animal behaviors. Drosophila melanogaster is an ...

SwarmSight: Real-time Tracking of Insect Antenna Movements and Proboscis Extension Reflex Using a Common Preparation and Conventional Hardware

Many scientifically and agriculturally important insects use antennae to detect the presence of volatile chemical compounds and extend their proboscis during feeding. The ability to rapidly obtain high-resolution measurements of natural antenna and proboscis movements and assess how they change in response to chemical, developmental, and genetic manipulations can aid the understanding of insect behavior. By extending our previous work on assessing aggregate insect swarm or animal group movements from natural and laboratory videos using the video analysis software SwarmSight, we developed a novel, free, and open-source software module, SwarmSight Appendage Tracking (SwarmSight.org) for frame-by-frame tracking of insect antenna and proboscis positions from conventional web camera videos using conventional computers. The software processes frames about 120 times faster than humans, performs at better than human accuracy, and, using 30 frames per second (fps) videos, can capture antennal dynamics up to 15 Hz. The software was used to track the antennal response of honey bees to two odors and found significant mean antennal retractions away from the odor source about 1 s after odor presentation. We observed antenna position density heat map cluster formation and cluster and mean angle dependence on odor concentration.

This protocol describes steps for using the novel software, SwarmSight, for frame-by-frame tracking of insect antenna and proboscis positions from conventional web camera videos using conventional computers. The free, open-source software processes frames about 120 times faster than humans and performs at better than human accuracy.

SwarmSight: Seguimiento en tiempo real de movimientos de insecto de antena y reflejo de extensión probóscide utilizando una preparación común y Hardware convencional

Many scientifically and agriculturally important insects use antennae to detect the presence of volatile chemical compounds and extend their proboscis ...

Robot controlado por insectos: una plataforma del robot móvil para evaluar la capacidad de seguimiento de Olor de un insecto

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