Published: September 2nd, 2016
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....
1. Study Animal
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 (.......
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.......
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....
|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
|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
|Bluetooth remote control to send the command to the operator laptop
|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
|8 cameras for a flight arena of 12.5 x 8 x 4 m
|Motion capture system
|12 cameras for a flight arena of 12.5 x 8 x 4 m
|3.7V, 10 mAh
|Retro reflective tape
|V92 reflective tape, silver color
|PFA-Insulated Silver Wire
|127 µm bare, 177.8 µm coated, 3 mm bare silver flame exposed at tips
|SMT Micro Header
|0.3 x 6 mm, bend to make a 3 mm long slider to secure the electrode into the PCB header.
|Secure the electrodes
|Immobilize the beetle
|Size 00; 0.3mm Rod diameter; 0.03 mm tip width; 38 mm Length
Make electrode guiding holes on cuticle
|Pattern #5; .05 X .01mm Tip Size; 110mm Length
Dissecting and implantation
|Vannas Micro Dissecting Spring Scissors; Straight; 3mm Cutting Edge; 0.1mm Tip Width; 3" Overall Length
Dissecting and implantation
|Potable soldering iron
|Melting beeswax and dental wax
|Light the entire flight arena with 30 panels (60 x 60 cm2). Each panel has 3 lamps.
14 W, 549 mm x 17 mm
Copyright © 2024 MyJoVE Corporation. All rights reserved