מערכת היברידית חרקים-מכונה: רדיו שליטה מרחוק של חיפושית עפה חופשי (<em> Mercynorrhina torquata</em>)
Summary
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.
Abstract
עלייתו של מכשירים אלקטרוניים דיגיטליים מאופשרת רדיו עוררה את השימוש מקליט לגירוי תוקפת אלחוטיים קטנים ללימוד התנהגות חרקים בזמן הטיסה. טכנולוגיה זו מאפשרת בניית מערכת היברידית חרק-אוטומטית באמצעות פלטפורמת חרק חייה המתוארת בפרוטוקול זה. יתר על כן, פרוטוקול זה מציג את תצורת מערכת פרוצדורות טיסה חינם להערכת התפקוד של שרירי התעופה ב חרק ומשוחרר. להדגמה, אנו ממוקדים sclerite השחי השלישיים (3Ax) השרירים לשלוט ולהשיג מפנה שמאלה או ימינה של חיפושית עפה. אלקטרודה חוט כסף דקה הושתלה על שריר 3Ax בכל צד של החיפושית. אלה חוברו התפוקות של תרמיל אלחוטי (כלומר, ממריץ חשמל תוקפת) רכוב על pronotum של החיפושית. השריר היה מגורה בטיסה חינם לסירוגין בצד גירוי (ימין או שמאל) או גיוון stimulatioתדירות n. החיפושית פנתה אל צד ipsilateral כאשר השריר היה מגורה והציג תגובה מדורגת אל בתדירות הולכת וגוברת. תהליך ההשתלה וכיול נפח של 3 מערכת המצלמה ללכוד תנועה ממדי צריך להתבצע בזהירות כדי למנוע נזק לשריר לאבד המסלול של הסמן, בהתאמה. שיטה זו היא מאוד מועילה ללמוד טיסת חרקים, כפי שהוא מסייע לחשוף את הפונקציות של שריר הטיסה של עניין טיסה חינם.
Introduction
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.
Protocol
Representative Results
Discussion
תהליך ההשתלה הוא חשוב, מאחר שהוא משפיע על מהימנותם של הניסוי. האלקטרודות צריך להיות מוכנס לתוך השריר בעומק של 3 מ"מ או פחות, תלוי בגודל של החיפושית (הימנעות ממגע עם שרירים באזור). אם אלקטרודות לגעת שרירים הסמוכים, פעולות מוטוריות רצויות והתנהגויות עלולות להתרחש בשל ?…
Offenlegungen
The authors have nothing to disclose.
Acknowledgements
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.
Materials
| Mecynorrhina torquata beetle | Kingdom of Beetle Taiwan | 10 g, 8 cm, pay load capacity is 30% of the body mass Aproval of importing and using by Agri-Food and Veterinary Authority of Singapore (AVA; HS code: 01069000, product code: ALV002). |
|
| Wireless backpack stimulator | Custom | TI CC2431 micocontroler The board is custom made based on the GINA board from Prof. Kris Pister’s lab. The layout of GINA board can be found at https://openwsn.atlassian.net/wiki/display/OW/GINA |
|
| Wii Remote control | Nintendo | Bluetooth remote control to send the command to the operator laptop | |
| BeetleCommander v1.8 | Custom. Maharbiz group at UC Berkeley and Sato group at NTU | Establish the wireless communication of the backpack and the operator laptop. Configure the stimulus parameters and log the positional data. Visualize the flight data. | |
| GINA base station | Kris Pister group at UC Berkeley | TI MSP430F2618 and AT86RF231 | |
| Motion capture system | VICON | T160 | 8 cameras for a flight arena of 12.5 x 8 x 4 m |
| Motion capture system | VICON | T40s | 12 cameras for a flight arena of 12.5 x 8 x 4 m |
| Micro battery | Fullriver | 201013HS10C | 3.7V, 10 mAh |
| Retro reflective tape | Reflexite | V92-1549-010150 | V92 reflective tape, silver color |
| PFA-Insulated Silver Wire | A-M systems | 786000 | 127 µm bare, 177.8 µm coated, 3 mm bare silver flame exposed at tips |
| SMT Micro Header | SAMTEC | FTSH-110-01-L-DV | 0.3 x 6 mm, bend to make a 3 mm long slider to secure the electrode into the PCB header. |
| Beeswax | Secure the electrodes | ||
| Dental Wax | Vertex | Immobilize the beetle | |
| Insect pin | ROBOZ | RS-6082-30 | Size 00; 0.3mm Rod diameter; 0.03 mm tip width; 38 mm Length Make electrode guiding holes on cuticle |
| Tweezers | DUMONT | RS-5015 | Pattern #5; .05 X .01mm Tip Size; 110mm Length Dissecting and implantation |
| Scissors | ROBOZ | RS-5620 | Vannas Micro Dissecting Spring Scissors; Straight; 3mm Cutting Edge; 0.1mm Tip Width; 3" Overall Length Dissecting and implantation |
| Potable soldering iron | DAIYO | DS241 | Reflow beeswax |
| Hotplate | CORNING | PC-400D | Melting beeswax and dental wax |
| Flourescent lamp | Philips | TL5 14W | Light the entire flight arena with 30 panels (60 x 60 cm2). Each panel has 3 lamps. 14 W, 549 mm x 17 mm |
Referenzen
- Kutsch, W., Schwarz, G., Fischer, H., Kautz, H. Wireless Transmission of Muscle Potentials During Free Flight of a Locust. J. Exp. Biol. 185 (1), 367-373 (1993).
- Fischer, H., Kautz, H., Kutsch, W. A Radiotelemetric 2-Channel Unit for Transmission of Muscle Potentials During Free Flight of the Desert Locust, Schistocerca Gregaria. J. Neurosci. Methods. 64 (1), 39-45 (1996).
- Ando, N., Shimoyama, I., Kanzaki, R. A Dual-Channel FM Transmitter for Acquisition of Flight Muscle Activities from the Freely Flying Hawkmoth, Agrius Convolvuli. J. Neurosci. Methods. 115 (2), 181-187 (2002).
- Sanchez, C. J., et al. Locomotion control of hybrid cockroach robots. J. R. Soc. Interface. 12 (105), (2015).
- Sato, H., et al. A cyborg beetle: insect flight control through an implantable, tetherless microsystem. , 164-167 (2008).
- Bozkurt, A., Gilmour, R. F., Lal, A. Balloon-Assisted Flight of Radio-Controlled Insect Biobots. IEEE Trans. Biomed. Eng. 56 (9), 2304-2307 (2009).
- Sato, H., et al. Remote Radio Control of Insect Flight. Front. Neurosci. 3, (2009).
- Daly, D. C., et al. A Pulsed UWB Receiver SoC for Insect Motion Control. IEEE J. Solid-State Circuits. 45 (1), 153-166 (2010).
- Maharbiz, M. M., Sato, H. Cyborg Beetles. Sci. Am. 303 (6), 94-99 (2010).
- Tsang, W. M., et al. Remote control of a cyborg moth using carbon nanotube-enhanced flexible neuroprosthetic probe. , 39-42 (2010).
- Hinterwirth, A. J., et al. Wireless Stimulation of Antennal Muscles in Freely Flying Hawkmoths Leads to Flight Path Changes. PloS ONE. 7 (12), (2012).
- Whitmire, E., Latif, T., Bozkurt, A. Kinect-based system for automated control of terrestrial insect biobots. , 1470-1473 (2013).
- Sato, H., et al. Deciphering the Role of a Coleopteran Steering Muscle via Free Flight Stimulation. Curr. Biol. 25 (6), 798-803 (2015).
- Erickson, J. C., Herrera, M., Bustamante, M., Shingiro, A., Bowen, T. Effective Stimulus Parameters for Directed Locomotion in Madagascar Hissing Cockroach Biobot. PLoS ONE. 10 (8), e0134348 (2015).
- Zhaolin, Y., et al. A preliminary study of motion control patterns for biorobotic spiders. , 128-132 (2014).
- Feng, C., Chao, Z., Hao Yu, C., Sato, H. Insect-machine hybrid robot: Insect walking control by sequential electrical stimulation of leg muscles. , 4576-4582 (2015).
- Cao, F., et al. A Biological Micro Actuator: Graded and Closed-Loop Control of Insect Leg Motion by Electrical Stimulation of Muscles. PLoS ONE. 9 (8), e105389 (2014).
- Zhao, H., et al. Neuromechanism Study of Insect-Machine Interface: Flight Control by Neural Electrical Stimulation. PLoS ONE. 9 (11), e113012 (2014).
- Tsang, W. M., et al. Flexible Split-Ring Electrode for Insect Flight Biasing Using Multisite Neural Stimulation. IEEE Trans. Biomed. Eng. 57 (7), 1757-1764 (2010).
- Barron, A. B. Anaesthetising Drosophila for behavioural studies. J. Insect Physiol. 46 (4), 439-442 (2000).
- Cooper, J. E. Anesthesia, Analgesia, and Euthanasia of Invertebrates. ILAR Journal. 52 (2), 196-204 (2011).
- Miller, T. A. . Insect neurophysiological techniques. , (2012).
- Leary, S., et al. . AVMA guidelines for the euthanasia of animals. , (2013).
- Heath, B., West, G., Heard, D., Caulkett, N. Mobile Inhalant Anesthesia Techniques. in Zoo Animal and Wildlife Immobilization and Anesthesia. , 75-80 (2008).
- Mischiati, M., et al. Internal models direct dragonfly interception steering. Nature. 517 (7534), 333-338 (2015).
- Kutsch, W., Berger, S., Kautz, H. Turning Manoeuvres in Free-Flying Locusts: Two-Channel Radio-Telemetric Transmission of Muscle Activity. J. Exp. Zoolog. Part A Comp. Exp. Biol. 299 (2), 139-150 (2003).
- Wang, H., Ando, N., Kanzaki, R. Active Control of Free Flight Manoeuvres in a Hawkmoth, Agrius Convolvuli. J. Exp. Biol. 211 (3), 423-432 (2008).
- Sato, H., Maharbiz, M. M. Recent developments in the remote radio control of insect flight. Front. Neurosci. 4, (2010).
- Tien Van, T., et al. Flight behavior of the rhinoceros beetle Trypoxylus dichotomus during electrical nerve stimulation. Bioinsp. Biomim. 7 (3), 036021 (2012).
- Sane, S. P., Dickinson, M. H. The control of flight force by a flapping wing: lift and drag production. J. Exp. Biol. 204 (15), 2607-2626 (2001).
- de Croon, G. C., et al. Design, aerodynamics and autonomy of the DelFly. Bioinsp. Biomim. 7 (2), 025003 (2012).
- Ma, K. Y., Chirarattananon, P., Fuller, S. B., Wood, R. J. Controlled Flight of a Biologically Inspired, Insect-Scale Robot. Science. 340 (6132), 603-607 (2013).
