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. Çalışma Hayvan <ol><li> Arka bireysel Mecynorrhina torquata böcekleri ahşap pelet yatak ile ayrı plastik kaplarda (6 cm, 8 g). </li><li> Her böceği şeker jöle (12 ml) 3 günde bir fincan besleyin. </li><li> sırasıyla 25 ° C ve% 60 balık yetiştirme oda sıcaklığı ve nem tutun. </li><li> ince tel elektrotlar implante önce her böceğin uçuş kapasitesini test edin. <ol><li> Yavaşça havaya bir böcek atmak. böceği 5 ardışık denemeler için daha uzun 10 saniye uçabilir ise böceği normal uçuş yetenekleri olduğu sonucuna varıldı ve sonraki uçuş deneyleri için kullanır. koyu hale getirmek için odadaki tüm ışıkları kapatın, böceği hatırlamak için. Bu uçuş sonlandırmak böceği neden olur. Not: Havaya salınan zaman bir böcek kendiliğinden uçup başlar. Böyle Şekil 1 (16 x 8 x 4 m gösterildiği gibi büyük bir kapalı bir odada uçuş deneyler daha iyidir <suBir hareket yakalama alanı ile p&gt; 3 12.5 x 8 x 4 m 3), havada açarken bir uçan böcek hareket çok hızlı (yaklaşık 3-5 m / sn) ve büyük arklar çizer olarak. </li></ol></li></ol> 2. Elektrot İmplantasyon <ol><li> 1 dakika 13,16,20-24 için CO 2 ile dolu bir plastik kapta koyarak böceği uyutmak. </li><li> 10 saniye sıcak suya batırarak diş balmumu yumuşatın. ahşap blok üzerinde anestezi böceği yerleştirin ve yumuşatılmış diş balmumu ile hareketsiz. Diş balmumu doğal soğutur ve birkaç dakika içinde katılaşır. </li><li> 25 mm uzunlukta kesilmiş yalıtımlı gümüş teller (127 mikron çıplak çapı, perfloroalkoksi ile kaplanmış 178 mikron çapında) implantasyon için ince tel elektrotlar kullanmak için. </li><li> Her bir hattın her iki ucundan da yalıtkan alev ile çıplak gümüş 3 mm Açığa. </li><li> Bir SMA oluşturmak için ince uçlu bir makas kullanarak böceğin manikür üst yüzeyini teşrihmetepisternum (Şekil 2c) yaklaşık 4 x 4 mm ll penceresi. Not: Şekil 2c gösterildiği gibi bir yumuşak kahverengi renkli manikür sonra, maruz kaldığı - e. 3AX kas yumuşak manikür altında yer almaktadır. </li><li> Pierce, iki deliği (Şekil 2d) ile 2 mm arasında bir mesafede olan bir böcek pin (boyut 00) kullanarak maruz kalan kahverengi kütikül iki delik açın. </li><li> dikkatli bir şekilde deliklerden (adım 2.4 hazırlanan bir aktif ve bir dönüş elektrotları dahil) iki kaynak teli yerleştirin ve 3 mm&#39;lik bir derinliğe her 3AX kas içine implant. </li><li> implante elektrotları Güvenli ve delikler üzerine eritilmiş balmumu bırakarak temas ve kısa devreyi önlemek için bir yerde tutun. Gerekirse, bir sıcak havya ucuyla balmumu dokunarak manikür üzerinde balmumu yeniden akar. balmumu hızla katılaşır ve implantasyon güçlendiriyor. Not: implantasyon doğru ise, pancar elitra kontrol etmek içinle elektrik stimülasyonu sırasında 3AX kas hareketini gözlemlemek için kaldırılabilir. </li></ol> 3. Kablosuz Sırt Çantası Meclisi Not: sırt çantası yerleşik radyo mikrodenetleyici 4 katmanlı FR-4 gemide (1.6 x 1.6 cm 2) oluşuyordu. sırt çantası, bir lityum polimer microbattery (3,7 V, 350 mg, 10 mAh) tarafından tahrik edildi. pil içeren sırt toplam kütlesi böceği (10 g vücut ağırlığı olan% 30) yük taşıma kapasitesine az 1.2 ± 0.26 g&#39;dır. sırt çantası kablosuz iletişimleri almak için önceden programlanmış ve iki çıkış kanalı vardı. <ol><li> Fosilden yüzeyini temizleyin çift taraflı bant kullanarak (manikür balmumu katmanı kaldırmak). Ardından, çift taraflı bant bir parça böceğinin Fosilden üzerinde sırt çantası takın. </li><li> sırt çıkışlarına yerleştirilen elektrotlar uçlarını bağlayın. </li><li> fo işaretleyici üretmek için microbattery etrafında yansıtıcı bant sarınr hareket yakalama kameraları tespit etmek için. </li><li> yansıtıcı bant hareket yakalama kameraları tarafından tespit edilebilir ve böylece çift taraflı bant bir parça kullanarak sırt çantası üstüne microbattery takın. </li></ol> 4. Kablosuz Kontrol Sistemi Not: Bu durumda, terim kablosuz kontrol sistemi uzaktan kumanda için bir alıcı içerir, özel uçuş kontrol yazılımı çalıştırmak için bir dizüstü bilgisayar, bir baz istasyonu, sırt çantası ve hareket yakalama sistemi. <ol><li> USB portları ile dizüstü bilgisayara uzaktan kumandanın baz istasyonu ve alıcıyı bağlayın. </li><li> Hareket yakalama sistemi açın ve bir Ethernet portu üzerinden dizüstü bilgisayara bağlayın. </li><li> Tamamen hareket yakalama alanı kapsayacak şekilde (motion capture sistemi satıcı şirket tarafından sağlanan) Kalibrasyon değnek sallayarak ses ayarı yapın. <ol><li> laptop masaüstünden hareket yakalama yazılımını açın. Tıklayın ve drag &quot;Kaynaklar&quot; paneli &quot;Sistem&quot; menüsündeki tüm kameraları seçin. </li><li> &quot;3D Perspektif&quot; menüsünü tıklayın ve kamera görünümüne geçmek için &quot;Kamera&quot; seçeneğini seçin. kalibrasyon ayarı göstermek için &quot;Araçlar&quot; panelinde &quot;Kamera&quot; sekmesine tıklayın. kameralardan gürültüyü ortadan kaldırmak için &quot;Kamera Maskeler oluşturma&quot; menüsünde &quot;Başlat&quot; düğmesini tıklayın ve gürültü mavi maskeli sonra sonra &quot;Dur&quot;. </li><li> Tıklayın ve &quot;Wand&quot; menüsünde ve &quot;Kamera&quot; sekmesinde &quot;L-Frame&quot; menüsünden &quot;5 Marker Wand &amp; L-Frame&quot; i seçin. 2,500 &quot;Wand Sayısı&quot; olarak ayarlayın, &quot;Kalibre Kameralar&quot; menüsünde &quot;Başlat&quot; a tıklayın ve tüm hareket yakalama uzayda kalibrasyon değnek dalga. asa sayısı 2.500 ulaştığında kalibrasyon işlemi durdurulur. </li><li> ( &quot;Araçlar&quot; panelinin &quot;Kamera&quot; sekmesinin altındaki) görüntü hatası daha yüksek 0,3 f ise kalibrasyon işlemi tekrarlayınveya herhangi bir kamera. ayarladıktan sonra hareket yakalama alanı ortasında yere değnek koymak ve hareket yakalama alanı kökenini belirlemek için &quot;Set Volume Origin&quot; menüsünde &quot;Başlat&quot; a tıklayın. </li></ol></li><li> Hareket yakalama uzayda bir kullanıcı tarafından salladı bir işaretleyici hareket yolunu kaydetmek ve işaretleyici algılanır ve izlenir olmadığını teyit etmek için bir kukla testi kullanılarak motion capture sistemi kapsama kontrol ediniz. işaretleyici sık algılama sırasında kesilirse kukla testi başarılı oluncaya kadar, hacim kalibrasyonunu tekrarlayın. <ol><li> &quot;Araçlar&quot; panelinde &quot;Yakalama&quot; sekmesine tıklayın ve daha sonra kendi yörüngesini kaydetmek için tüm hareket yakalama uzayda örnek işaretleyici sallayarak önce &quot;Yakalama&quot; menüsünde &quot;Başlat&quot;. </li><li> Kayıttan sonra, üzerine tıklayın marker konumlarını yeniden ve kayıt kalitesini kontrol etmek için &quot;Yeniden İnşa boru hattı çalıştırır&quot;. </li></ol></li><li> microb terminallerini bağlayınattery sırt çantası güç pinlerine (adım 3.4 sırt çantası takılı). </li><li> dizüstü bilgisayar ve özel uçuş kontrol yazılımını kullanarak sırt çantası arasında kablosuz iletişimi test. yazılım üzerinde &quot;Başlat&quot; komutunu tıklayın ve görüntülenen bağlantı durumunu kontrol edin. </li></ol> 5. Serbest Uçuş Denemesi <ol><li> 16 x 8 x 4 m 3 boyutlarında bir uçuş arenada ücretsiz bir uçuş deneyi yürütmek. </li><li> Giriş uçuş kontrol yazılımı (voltaj, darbe genişliği, frekans ve stimülasyon süresi) uygun parametreler. Not: gösteri için, 3 msn, 3 V darbe genişliği gerilimini sabit ve stimülasyon süresi 1 sn ve 60 ila 100 Hz frekansı değişiyordu. <ol><li> &quot;Gerilim&quot; kutusuna 3 V yazılım ekranında, tip 3, &quot;Uyarım Süresi&quot; kutusuna 1.000 msn, &quot;Genişlik Pulse&quot; kutusuna 3 ms için 3 1.000, ve Hz istenen frekansta &quot; frekans &quot;kutu on komut penceresi. </li></ol></li><li> o uçuş arenada içinde serbestçe uçmak için izin havaya sırt çantası monte böceği bırakın. böceği hareket yakalama alanı girdiğinde manuel stimülasyon tetikler. Basın uzaktan kumanda üzerindeki uygun komut düğmesi (Sol veya Sağ) böceğinin sağ veya sol taraftaki hedef kas uyarmak için. Not: düğmesine basıldığında, dizüstü bilgisayar üzerinde çalışan uçuş kontrol yazılımı komutu oluşturur ve sırt çantası gönderir. sırt çantası sonra (sağ veya sol tarafta) faiz kas elektriksel uyaran verir. </li><li> stimülasyon sırasında gerçek zamanlı olarak böceğin tepkisini gözlemleyin ve 3D grafik yazılımı kullanarak veri yeniden. <ol><li> &quot;Böcek Display&quot; penceresinin veri listesinde kayıtlı çalışmaların birini seçin ve analiz klasörüne bu araştırmanın verileri kopyalamak ve 3D grafik modülü çalıştırmak için &quot;İhracat Panda&quot; tıklayın. </li><li> Basın &quot;N&quot;Klavye kaydedilen yörünge ile uyaran sinyalini birleştirmek. Basın Vurgulanan uyarım dönemleri ile böceği yörüngesini gösterir. </li></ol></li></ol>

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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.

bu deney güvenilirliğini etkileyen olarak implantasyon süreci önemlidir. Elektrotlar 3 mm böceği (yakın kas temas kaçınarak) boyutuna bağlı olarak ya da daha az bir derinlikte de kas içine yerleştirilmelidir. Elektrotlar yakın kasları dokunursanız, istenmeyen motorlu eylem ve davranışlar yakın kasların kasılması nedeniyle oluşabilir. iki elektrot iyi hiçbir kısa devre meydana sağlamak için aynı hizada olmalıdır. erime ve bir havya kullanarak balmumu yeniden düzenlenerek zaman Deneyciler di...

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)

radyo özellikli dijital elektronik cihazların yükselişi de uçuş böcek davranışlarını incelemek için küçük kablosuz nöromüsküler kaydediciler ve uyarıcıların kullanımı teşvik etti. Bu teknoloji bu protokolü açıklanan yaşayan bir böcek platformu kullanarak bir böcek-makine hibrid sisteminin geliştirilmesini sağlar. Ayrıca, bu protokol ve sistem konfigürasyonu zincirlenmemiş böcek uçuş kaslarının fonksiyonunu değerlendirmek için ücretsiz bir uçuş deneysel prosedürler sunar. gösteri için, kontrol ve uçan böcek sol veya sağ dönüm elde etmek için üçüncü aksiller sclerite (3AX) kas hedef. Ince gümüş tel elektrot böceği her tarafta 3AX kası üzerine yerleştirildi. Bunlar kablosuz sırt çantası çıkışlarına bağlı idi (yani, bir nöromusküler elektrik uyarıcısı) böceği Fosilden üzerine monte edilmiş. kas (sol veya sağ) stimülasyon tarafını alternatif veya stimulatio değiştirilerek serbest uçuşta uyarıldıN frekansı. kas uyarılır ve giderek artan sıklıkta bir kademeli tepki sergiledi zaman böceği ipsilateral tarafa döndü. implantasyon süreci ve 3 boyutlu hareket yakalama kamera sistemi hacim kalibrasyonu, sırasıyla kas zarar ve işaretleyici izini kaybetmemek için dikkatli yürütülen gerekir. serbest uçuş ilgi uçuş kas fonksiyonlarını ortaya çıkarmak için yardımcı olur Bu yöntem, böcek uçuşu incelemek için son derece faydalıdır.

Elektrot yerleştirme işlemi Şekil 2&#39;de sunulmuştur ince gümüş tel elektrotlar kas (Şekil 2d - e) yumuşak manikür deldi küçük deliklerden böceği 3AX kasına implante edildi.. Bu yumuşak manikür metepisternum ön kısmını çıkardıktan sonra sadece basalar kas apodema üzerinde bulunan (Şekil 2d - c). Elektrotlar daha sonra balmumu (Şekil 2f) kullanılarak güvenc...

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.

Böcek-makine Hibrid Sistem: Serbest Uçan Beetle Uzaktan Radyo Kontrol (<em&gt; Mercynorrhina torquata</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 Görüntüleme.  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.

Böcek Uçuş Behavior Monitoring için erken Metamorfik Yerleştirme Teknoloji

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

Davranış

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.

Yetiştirme ve Gizle böcek RNA aracılı gen demonte çift sarmallı,<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 ...

Genetik

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.

Tork Metre Drosophila Edimsel Öğrenme

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 ...

Biyoloji

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.

Ambrosia Beetle, Platypus Quercivorus (Murayama), ucuz, küçük ve kolayca inşa uçuş Mill kullanarak uçuş yeteneği ölçme

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

Çevre

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.

Batı Mısır Rootworm, Diabrotica virgifera (LeConte) Uçuş Eğilimi ve Performansını Ölçmek için Flight Mills kullanma

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.

Serbestçe Böcekler Yürüyüş Beyin Ektraselüler Tel Tetrode Kaydı

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

Nörobilim

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.

Bağlı Böcek Uçuşunun Çalışması İçin Gelişmiş Bir Uçuş Değirmeni İnşa Etmek

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

Mühendislik

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.

Koku Aramalar için Otonom Robotlar Böcek elektroantenogram Sensörleri kullanarak

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.

Radyo Frekansı ile Tanımlama ve Verimli Hareket duyarlı video Bumblebees tarafından karşılıksız seçim davranışının Kayıt otomatikleştirin

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.

Böcekler Tethered Flight Araştırmaları Basit Uçuş Mill

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

Böcek-makine Hibrid Sistem: Serbest Uçan Beetle Uzaktan Radyo Kontrol (

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