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. animal Experimental <ol><li> Prepare uma caixa de plástico para manter a pupas de silkmoths do sexo masculino (B. mori) até à sua eclosão. Toalhas de papel colocadas na parte inferior e pedaços de papelão ao redor da parede interna da caixa (Figura 1A). Nota: Os pedaços de papelão são necessários para as mariposas adultas para segurar enquanto estende suas asas durante a eclosão (Figura 1A). </li><li> Coloque silkmoth masculino (Bombyx mor i) pupas na caixa e mantê-los em uma incubadora até a eclosão sob um 16-hr: Luz 8 horas: ciclo escuro a 25 ° C. NOTA: O macho e pupas do sexo feminino podem ser discriminados pelas marcações do sexo no abdômen (Figura 1B). </li><li> Recolha mariposas machos adultos após a eclosão e movê-los para uma nova caixa. </li><li> Manter as traças adultas numa incubadora sob uma 16-h: luz 8-h: ciclo escuro e diminuir a temperatura a 15 ° C para reduzir a sua actividade antes da experiência. </li></ol> 2. Amarrar um Silkmoth <ol><li> Fabricação de um anexo para o tethering (Figura 2A) Nota: O acessório é constituído por um fio de cobre com uma tira de uma folha fina de plástico, na sua ponta. Isto assegura o movimento dorso-ventral do tórax durante a marcha (Figura 2B). <ol><li> Prepara-se uma tira de uma folha fina de plástico, 2 × 40 mm (espessura: 0,1 mm), e dobrá-lo no meio. </li><li> Anexar a tira dobrada à ponta de um fio de cobre com um adesivo. </li><li> Dobre a ponta da tira dobrada, onde o tórax de um silkmoth está ligado. </li></ol></li><li> Use mariposas adultas (2-8 dias) durante o período de luz para o experimento. Nota: A sensibilidade para a feromona depende fortemente do relógio circadiano 18. Porque B. mori é uma mariposa diurna, o experimento deve ser realizado durante o período de luz. </li><li> Remova cuidadosamente todas as escalas na dorSal tórax (mesonoto) usando um pedaço de tecido húmido (ou haste de algodão) e expor a cutícula do mesonoto (Figura 2C). </li><li> Cole um adesivo sobre a tira de plástico na fixação e na superfície do mesonoto exposta com uma chave de fendas pequena e esperar 5-10 min, até que o adesivo deixa de ser viscoso. Nota: O adesivo não deve tocar a dobradiça asa ou o tegulae asa anterior (Figura 2C). </li><li> Unir o mesonoto ao anexo. </li><li> Mantenha a traça amarrado antes de colocá-lo dentro do cockpit do robô. Segure o acessório em um carrinho e colocar um pedaço de papel por baixo das pernas para descansar a mariposa. </li></ol> 3. Robô controlado-Insecto <ol><li> Projetar o hardware do robô controlado por inseto com base em trabalhos anteriores 16,17,19. Nota: O robô controlado por insetos consiste em uma esteira apoiado pelo ar com um sensor de rato óptico para Capture o inseto locomoção, à base de AVR placas de microcontroladores custom-built para processamento e controle motor, e dois motores DC sem escovas (Figuras 3 e 4). O robô pode ser executado com base na rotação bola com precisão de 96% ou superior, dentro de um tempo de atraso de 200 ms. Ele também garante a mobilidade de máxima velocidade de avanço (24.8 mm / s) e velocidade angular (96,3 ° / seg) do silkmoth durante feromônio comportamento de rastreamento 16. O fluxo de ar da esteira (Figura 5A) e sistema de entrega odor (Figura 5B) são projetados para a traça a bordo de caminhar sem problemas sobre a bola e adquirir um odor por duas antenas. A entrada de ar e do fluxo do canal do tapete rolante é separado dos do sistema de libertação de odores, para evitar a contaminação da feromona. </li><li> Projetar o software para os microcontroladores de bordo baseados em trabalhos anteriores 16. Nota: O microcontrolador calcula a bordo tele movimentos do robô da locomoção inseto medido com um sensor óptico (rotacional, Δ x; translacional, Δ Y; Figura 6). A distância de deslocamento (Δ L) e virar ângulo (Δθ) por unidade de tempo do robô são calculados com base na distância de percurso de cada uma das rodas (esquerda, Δ L L; direito, Δ L R), tais como Δ G = (Δ L L L + Δ R) / 2 e Δθ = (Δ L L - L Δ R) / roda de D, em que D roda é a distância entre as duas rodas (120 mm). Δ L L e Δ L R são ainda descritos como Δ G G = x Δ L, L + Δ G Y, L e Δ L R = X Δ G, Y, R, em que X Δ L, L &lt;Δ L R +/ sub&gt; e x Δ L, R são as distâncias das rodas nos lados esquerdo e direito controladas por Δ x, e Δ L y, L e Δ L y, R são as empresas controladas por Δ y. Idealmente, Δ x G, L e X Δ L, R são descritos como X Δ L, L = X -Δ L, R = G Δ x (D roda / bola D), e Δ G Y, L e L Y Δ , R são descritos como Δ G Y, L = Δ L Y, R = G Δ Y, em que G é o ganho do motor e esfera D é o diâmetro da bola (50 mm). Na prática, o ganho do motor é definido independentemente por cada um dos lados (esquerdo ou direito da roda) e por cada direcção (para a frente ou para trás de rotação) de modo a calibrar o movimento do robô. Os ganhos independentes permitem ainda mais para oconfiguração assimétrica de rotação do motor para gerar um viés de rotação do robô (ver passo 6.1). </li><li> Lava-se a superfície de uma bola branca Poliestireno (de massa: aproximadamente 2 g; diâmetro: 50 mm) com água para remover quaisquer possíveis pistas olfactivas ou visuais. Nota: A superfície de uma nova bola deve ser desbastada com lixa fina, tais como P400, o que assegura o aperto das pernas sobre a bola. </li><li> Ligue o ventilador do ventilador que fornece ar às 9 V para a esteira e flutua a bola (Figura 5A). Observar a esfera flutuante de aproximadamente 2 mm a partir do fundo do copo. </li><li> Usando um parafuso, fixe o fio de cobre do anexo com a traça (consulte a etapa 2) para um dispositivo elétrico na cabine do robô (ver Figura 3 inserção). Certifique-se de que a posição das pernas do meio está no centro da esfera (Figura 7A). </li><li> Ajuste a posição vertical da fixação para permitir a traça a andar normalmente no btodos. Manter a bola na mesma altura antes e depois de anexar a traça (Figura 7B). Nota: Uma posição muito menor do acessório adiciona pressão sobre a traça e elicia uma curta para trás, para resistir à pressão (Figura 7C), ao passo que uma posição alta demais provoca andar instável e falhas do sensor devido a alterações na posição vertical de a bola (Figura 7D). Para verificar o comportamento de caminhada normal, um single-inchado estímulo feromônio é usado para disparar andando na traça (para o estímulo feromônio, consulte o passo 4). Note-se que o estímulo de teste deve ser mínimo, porque a exposição anterior para bombykol habitua silkmoths e diminui a sua sensibilidade (Matsuyama e Kanzaki, dados não publicados). </li></ol> 4. Preparação Fonte Odor Nota: Masculino B. mori são sensíveis ao componente principal da feromona sexual feminina da mesma espécie (bombykol: (E, Z) -10,12-hexadecadien-1-ol) <s-se&gt; 20. Qualquer contaminação do equipamento experimental com bombykol provoca o comportamento de rastreamento de odor e afecta a capacidade de resposta da traça. <ol><li> Gota de 10 ul da solução bombykol dissolvido em n-hexano (200 ng / ul) sobre um pedaço de papel de filtro (cerca de 10 mm × 10 mm). A quantidade de bombykol cada pedaço de papel de filtro é de 2.000 ng. Nota: Para verificar o comportamento normal curta da mariposa, prepare um cartucho de feromônio estímulo nesta etapa. O cartucho é uma pipeta de Pasteur de vidro com um pedaço de papel de filtro contendo 2000 ng de bombykol. Empurrando um bulbo incha o bombykol ar que contém. </li></ol> 5. Odor Fonte Localization Experiment <ol><li> Ligue o ventilador de um túnel de vento ao ar do tipo de puxar (1.800 × 900 mm × 300 mm, L × W × H; Figura 8) e definir a velocidade do vento a 0,7 m / s. Certifique-se de que a temperatura é superior a 20 ° C. </li><li> Defina a fonte de odor (o pIECE de papel de filtro contendo bombykol) a montante do túnel de vento. Nota: A largura pluma deve ser confirmado antes da experiência usando TiCl4 17,19. </li><li> Ligue a placa do microcontrolador do robô e estabelecer uma conexão serial a um PC via Bluetooth. </li><li> Lançar um programa Java feito por encomenda chamado de &quot;sinais vitais&quot;, que fornece uma interface entre o PC eo robô. Nota: A janela principal inclui botões para enviar comandos para o robô, janelas de texto para exibir a entrada e saída de comunicação serial, e pequenas caixas para configurar os parâmetros. Os comandos subseqüentes são enviadas clicando nos botões correspondentes no programa, exceto para captura de vídeo. </li><li> Clique no &quot;sobre o dispositivo&quot; botão para confirmar a conexão enviando um comando para o robô através da porta COM especificado e verifique se uma mensagem é devolvido pelo robô. </li><li> Clique no botão &quot;memory apagar &quot;botão para apagar dados de locomoção anteriores deixadas na memória flash onboard. </li><li> Clique no botão &quot;drivemode1&quot; para enviar os ganhos motores padrão para o robô. Nota: As manipulações dos ganhos motor e o tempo de atraso entre a locomoção de insetos e movimento do robô são aplicadas após esta etapa (veja as etapas 6.1 e 6.3, figura 9). </li><li> Clique no botão &quot;não conduza&quot; para enviar um comando para imobilizar o robô até que a experiência começa. </li><li> Coloque o robô em uma posição de partida (600 mm a jusante da fonte de odor) e ligue o interruptor da placa de excitador do motor. </li><li> Pressione o botão de gravação da câmera de vídeo para iniciar a captura de vídeo. </li><li> Clique no botão &quot;start rec&quot; para enviar um comando de partida para iniciar o robô com uma gravação simultânea da rotação bola na memória flash onboard. Observe que o robô começa a se mover e rastreia a pluma de odor. </li><li> Clique no&quot;STOP REC&quot; e &quot;não dirija&quot; botões para enviar comandos para parar tanto o movimento do robô ea gravação se o robô localiza a fonte de odor. </li><li> Pressione o botão de gravação da câmera de vídeo para parar a captura de vídeo. </li><li> Baixar os dados gravados de locomoção da memória flash integrada ao computador através de uma conexão serial. Feche o programa. </li></ol> 6. A manipulação do robô controlado por Insect Nota: O tempo de cada manipulação é indicado na Figura 9. <ol><li> Manipulação de ganhos motores Nota: Esta manipulação altera a velocidade de translação e de rotação do robô. Ganhos motores assimétricos gerar um viés de torneamento, que pode ser usado para investigar como insectos compensar a polarização 17. <ol><li> Definir os ganhos de rotação para a frente e rotação para trás do motor de cada lado 17 (Figura 6B) editando tharquivo e de configuração chamado &quot;param2.txt&quot; usando um editor de texto. </li><li> Clique no botão &quot;param2 set&quot; para ler o arquivo de configuração editado no programa de software. Em seguida, clique no botão &quot;drivemode2&quot; para enviar os ganhos manipulados para o robô. </li></ol></li><li> Inversão da saída do motor Nota: Esta manipulação fornece uma condição similar à inversão de entrada olfactiva bilateral (ver passo 6.4) e pode ser usado para investigar o significado do olfacto bilateral. No entanto, a inversão da saída do motor também inverte movimento visual auto-indução de uma traça a bordo. O impacto da entrada visual de auto-induzida invertido pode ser avaliada através de uma comparação com a entrada invertida 19 olfactiva. <ol><li> Inverter o controle motor bilateral através do cruzamento dos cabos de controlo para cada motor. </li></ol></li><li> Manipulação do tempo de atraso entre a locomoção e de insectos movimento do robô. Nota: Esta manipulaçãopermite a investigação do período aceitável de tempo gasto no processamento sensorial-motor para o odor-tracking robótico. O microcontrolador armazena os dados de locomoção em um buffer de memória e, em seguida, processa-lo após o intervalo de tempo especificado. Note-se que o robot tem um atraso de tempo interna máxima de 200 ms; Portanto, o retardo de tempo real deve ser o intervalo de tempo especificado, acrescidos de 200 mseg 16,17. <ol><li> Introduza um número (0-10) em uma pequena caixa da janela principal para especificar um tempo de atraso de 0-1,000 ms em passos de 100 ms. </li><li> Clique no botão &quot;definir o atraso&quot; para aplicar o tempo de atraso. </li></ol></li><li> Manipulação da entrada olfactiva. Nota: Esta manipulação pode ser utilizada para investigar o significado de entrada olfactiva bilateral. A direção onda de silkmoths é tendenciosa no lado de maior concentração 22. <ol><li> Mudar a lacuna entre as pontas de tubo de sucção ou inverter as suas posições para alterar odiferença na concentração de odor adquirida por cada antena. </li></ol></li><li> Manipulação da informação visual Nota: Esta manipulação é investigar o papel da entrada visual para odor-tracking. <ol><li> Cobrir o dossel com um papel branco que obstrui 105 ° e 90 ° do campo visual horizontal e vertical da traça a bordo, respectivamente. </li></ol></li></ol>

The authors have nothing to disclose.

Os pontos mais importantes para o controle bem sucedido do robô por um silkmoth estão deixando a traça caminhar sem problemas sobre a bola apoiado pelo ar ea forma estável medir a rotação de bola. Por isso, amarrar o silkmoth e montá-lo sobre a bola na posição apropriada são os passos críticos neste protocolo. adesão inadequada da mariposa para a fixação ou posicionamento inadequado da mariposa na bola irá causar pressão antinatural sobre ele, o que perturba o seu comportamento marcha normal e / ou provo...

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.

Apresentamos aqui as características básicas do robô controlado por insetos necessário para a localização bem-sucedida de uma fonte de odor. A comparação entre o robot e silkmoths, a eficácia do sistema de entrega de odor, e o significado de olfactivo bilateral exacta e entradas visuais são examinados. A comparação dos comportamentos de odor-rastreamento entre traças livremente a pé e o robot controlado por insec...

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

A capacidade para localizar uma fonte de odor é necessária para a sobrevivência de insectos e espera-se que seja aplicável aos odores-rastreamento artificial. O robô controlado por insetos é accionada por um silkmoth real e nos permite avaliar a capacidade de rastreamento de odor de insetos através de uma plataforma robótica.

Robô controlado por insetos: uma plataforma móvel Robot avaliar a capacidade Odor-tracking de um inseto

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

Research

JoVE Journal

Neuroscience

14.6K visualizações.  The University of Tokyo.  A capacidade para localizar uma fonte de odor é necessária para a sobrevivência de insectos e espera-se que seja aplicável aos odores-rastreamento artificial. O robô controlado por insetos é accionada por um silkmoth real e nos permite avaliar a capacidade de rastreamento de odor de insetos através de uma plataforma robótica. 

Vídeo: 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 resolução de Medição do Comportamento Odor-Driven em larvas de Drosophila

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

Biologia

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.

Visualmente Rastreamento Odor Mediated Durante o Voo em 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.

Um túnel de vento para Odor mediada insetos ensaios comportamentais

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 ...

Comportamento

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.

Vôo Detecção e Classificação Insect com sensores baratos

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

Bioengenharia

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.

Ensaio Escolha Vertical labirinto em T para a resposta de Artrópodes para odores

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

Neurociência

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.

Usando insetos eletroantenogramas Sensores em robôs autônomos para Pesquisas 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 detecção de odor baseado em eletroantennografia usando antenas de seda para localização de fonte de odor

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

Engenharia

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 resolução Quantificação do Comportamento Odor-guiada em<em&gt; Drosophila melanogaster</em&gt; Usando a<em&gt; Flywalk</em&gt; Paradigm

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.

Comportamentos olfativos ensaiadas pelo monitoramento do computador de<em&gt; Drosophila</em&gt; Em um olfatômetro de quatro quadrantes

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: Acompanhamento em tempo real dos movimentos de antena do inseto e reflexo de extensão Proboscis usando uma preparação comum e Hardware convencional

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

Robô controlado por insetos: uma plataforma móvel Robot avaliar a capacidade Odor-tracking de um inseto

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