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Summary

Abstract

Introduction

Protocol

Representative Results

Discussion

Acknowledgements

Materials

References

Neuroscience

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

Published: September 2nd, 2016

DOI:

10.3791/54260

1School of Mechanical and Aerospace Engineering, 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.

The rise of radio-enabled digital electronic devices has prompted the use of small wireless neuromuscular recorders and stimulators for studying in-flight insect behavior. This technology enables the development of an insect-machine hybrid system using a living insect platform described in this protocol. Moreover, this protocol presents the system configuration and free flight experimental procedures for evaluating the function of the flight muscles in an untethered insect. For demonstration, we targeted the third axillary sclerite (3Ax) muscle to control and achieve left or right turning of a flying beetle. A thin silver wire electrode was implanted on the 3Ax muscle on each side of the beetle. These were connected to the outputs of a wireless backpack (i.e., a neuromuscular electrical stimulator) mounted on the pronotum of the beetle. The muscle was stimulated in free flight by alternating the stimulation side (left or right) or varying the stimulation frequency. The beetle turned to the ipsilateral side when the muscle was stimulated and exhibited a graded response to an increasing frequency. The implantation process and volume calibration of the 3 dimensional motion capture camera system need to be carried out with care to avoid damaging the muscle and losing track of the marker, respectively. This method is highly beneficial to study insect flight, as it helps to reveal the functions of the flight muscle of interest in free flight.

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 technolo....

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1. Study Animal

  1. Rear individual Mecynorrhina torquata beetles (6 cm, 8 g) in separate plastic containers with wood pellet bedding.
  2. Feed each beetle a cup of sugar jelly (12 ml) every 3 days.
  3. Keep the temperature and humidity of the rearing room at 25 °C and 60%, respectively.
  4. Test the flight capability of each beetle before implanting thin wire electrodes.
    1. Gently throw a beetle into the air. If the beetle can fly for longer than 10 sec for 5 consecutive trials, c.......

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The electrode implantation procedure is presented in Figure 2. Thin silver wire electrodes were implanted into the 3Ax muscle of the beetle through small holes pierced on the soft cuticle on the muscle (Figures 2d-e). This soft cuticle is found just above the apodema of the basalar muscle after removing the anterior part of the metepisternum (Figures 2d-c). The electrodes were then secured using beeswax (.......

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The implantation process is important, as it affects the reliability of the experiment. The electrodes should be inserted into the muscle at a depth of 3 mm or less depending on the size of the beetle (avoiding contact with nearby muscles). If the electrodes touch the nearby muscles, undesirable motor actions and behaviors may occur owing to the contraction of nearby muscles. The two electrodes should be well aligned to ensure that no short circuits occur. When melting and reflowing beeswax using a soldering iron, the ex.......

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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 a....

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Name Company Catalog Number Comments
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 

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