The authors would like to thank the George Washington University Facilitating Fund for financial support of the project. Mr. Patel is grateful the George Washington University School of Engineering and Applied Science Summer Undergraduate Program in Engineering Research and the Undergraduate Research award for financial support. Finally, we are grateful to the GWU Center for Biomemetics and Bioinspired Engineering (COBRE) for use of facilities controlled by the center.

1. Digitize a Specimen of a Sea Lion Foreflipper
<ol>
	<li>Scan a specimen of a Sea lion foreflipper.
	<ol>
		<li>Obtain a specimen of a sea lion flipper from a deceased individual (Figure 1a). 
		NOTE: In our case, they were obtained from the Smithsonian Zoological Park in Washington, D.C.</li>
		<li>Hang the foreflipper vertically from its base (where the foreflipper attaches to the animal&#39;s body). This both allows the flipper to be straight when scanned, and exposes the entire surface for scanning.</li>
		<li>Scan flipper using a high-resolution structured light scanner, with an accuracy of approximately 0.5 mm, and error of approximately 0.1 mm (Figure 1b).</li>
	</ol>
	</li>
	<li>Import the point cloud into CAD software and render it as a surface. To do this, click &#39;Open&#39; and select the desired .obj file. Click on &#39;Import&#39; to import the file into the CAD software.</li>
	<li>Manipulate the resulting point cloud using a computer-aided design (CAD) software by clicking on &#39;Extruded cut&#39; and cutting out the flesh part (unwanted part) of the scan. Next, click on &#39;Scale&#39; to obtain the appropriate scaling for the robotic flipper (68% of the full size). Inspect flipper for sufficient detail capture by comparing to the original specimen (Figure 2).</li>
	<li>Create the mold around the flipper.
	<ol>
		<li>In a CAD software, use the flipper surfaces to form a mold by creating a surrounding volume around the flipper surface. Do this by extruding a rectangular block by clicking on &#39;Sketch&#39; to draw a rectangle and then extruding it to more than the height of the flipper to completely encompass it.</li>
		<li>Click on &#39;Assembly&#39; and import both parts (flipper and rectangular block) into the working area. Click on &#39;Mate&#39; and make the front and top plane of both the flipper and mold as coincident. This automatically places the flipper inside the mold.</li>
		<li>Select the mold from the design tree and click &#39;Edit Part&#39;. Once the part is selected, click on &#39;Insert&#62;Features&#62;Cavity&#39; to make a cavity of the flipper inside the mold. Sketch a line at the center of the rectangular mold and click on &#39;Split&#39; to form two parts of the same mold.</li>
		<li>Click on &#39;Cut Part&#39; to separate the surrounding volume into two parts for easy flipper extraction. Insert cavities and pegs on each half of the volume and save it as part one and two of the flipper mold (Figure 3).</li>
		<li>Convert the &#39;.SLDRPT&#39; files of the mold to &#39;.STL&#39;. Import these files to the proprietary software of the 3D printer and click on &#39;Print&#39; to generate the 3D printed mold.</li>
	</ol>
	</li>
</ol>
2. Design the Bone Structure
<ol>
	<li>Open the digital foreflipper in a CAD software and obtain an image of the Sea lion foreflipper bone structure for reference (such as Figure 1 in English, 197719).</li>
	<li>Design three different pieces that mimic the bone structure that will fit inside the digital model of the foreflipper. Throughout this procedure, &#39;base&#39; refers to the end of a part closer to the base of the foreflipper and &#39;tip&#39; refers to the end of the part closer to the tip of the foreflipper.
	<ol>
		<li>Base Piece
		<ol>
			<li>Make the length of this piece proportional to the distance between the shoulder joint and the wrist of the Sea lion flipper (measurements are obtained using measuring tape). Do this using a CAD software by clicking on &#39;Sketch&#39; and designing the shape of the base piece (Figure 4).</li>
			<li>Add knuckles at both ends of the part by clicking on &#39;Sketch&#39; and drawing two circles. Click on &#39;Boss Extrude&#39; to extrude the desired length from the plane of the base piece. Click on the sketch of the smaller circle to cut into the extrude by clicking &#39;Cut Extrude&#39; to make room for the shaft. To strengthen this joint, click on &#39;Fillet&#39; to smoothen the sharp joints. 
			NOTE: The dimensions of the circles depend on the size of the shaft to be used during mounting the flipper on top of the water flume. In our case, the diameter of the smaller circle is 0.5 inches and the bigger circle is 1 inches. The base end will be sitting outside the flipper skin geometry, so the size of the knuckles does not fall under the constraints of the skin.</li>
		</ol>
		</li>
		<li>Middle Piece
		<ol>
			<li>Make the length of this piece proportional to the distance between the wrist joint and the knuckle joint of a Sea lion. Do this by clicking on &#39;Sketch&#39; and sketching the desired shape (as shown in Figure 4b) on a plane. Once the geometry is designed, click on &#39;Extrude&#39; to get the basic three-dimensional shape of the middle piece. Input the extruded length as 0.1650 inches. 
			NOTE: The desired shape of the middle piece in our experiment is a trapezoid with a height of 2.25 inches and the length of the two bases as 1.625 and 0.850 inches respectively.</li>
			<li>Add knuckles on both ends. Do this as described in step 2.2.1.2. The diameter of the extruded cut is 0.125 inches. Connect the knuckles on the base end to the tip end of the base piece with an axel to form a hinge representing the wrist joint. 
			NOTE: The knuckles need to fit inside the volume of the foreflipper, so design accordingly.</li>
			<li>Add a tower approximately 1 cm in height to the tip end of the piece on both sides.
			<ol>
				<li>To add a tower, click on &#39;Sketch&#39; and sketch a rectangle on the base of the model. Extrude the sketch by selecting the sketch and clicking on &#39;Boss Extrude&#39;. The thickness of the tower in this particular case is 0.165 inches.</li>
				<li>Click on &#39;Fillet&#39; and select the model and one edge of the extruded tower. This strengthens the sharp joint where the tower and the base of the middle piece are connected. It is okay if the tower protrudes from the geometry of the skin. The tower should be thick enough to withstand the forces generated during a flipper clap. See Figure 4 for reference.</li>
			</ol>
			</li>
		</ol>
		</li>
		<li>Tip Piece
		<ol>
			<li>Make the length of this piece proportional to the distance between the knuckle joint and the tip of the longest finger bone of a Sea lion. Do this by clicking on &#39;Sketch&#39; and sketching a desired shape on a plane. Once the geometry is designed, click on extrude to get the basic three-dimensional shape of the tip piece.</li>
			<li>Add knuckles on both ends. Do this as described in step 2.2.1.2. The diameter of the extruded cut should be equal to the diameter of the axle, which in this experiment is 0.125 inches. The knuckles on the base end will be connected to the tip end of the middle piece with an axle to form a hinge representing the knuckle joint. The geometry of these knuckles needs to fit inside the geometry of the foreflipper skin, so design accordingly.</li>
			<li>Add a tower approximately 1 cm in height to the base end of the piece on both sides. Do this described in step 2.2.2.3. The thickness of the tower in this particular case is 0.165 inches. It is okay if the tower protrudes from the geometry of the skin. The tower should be thick enough to withstand the forces generated during a flipper clap. See Figure 5 for reference.</li>
		</ol>
		</li>
	</ol>
	</li>
</ol>
3. Creating a Flipper
<ol>
	<li>3D print the skeleton (base, middle and tip pieces) of the flipper. Convert the &#39;.SLDRPT&#39; file from CAD to &#39;.STL&#39; and import it into the printer&#39;s proprietary software and click &#39;Print&#39;. 
	NOTE: The printing instructions are different for each printer.
	<ol>
		<li>Reinforce the knuckles of the middle and tip piece with an adhesive (epoxy) and carbon threads. To do this, cut carbon threads of length 0.750 inches. Apply adhesive to the 3D printed bone structures and lay the threads over the knuckles. It is not necessary to reinforce the large knuckles on the base piece (Figure 5a).</li>
		<li>Drill holes at the bottom of each tower the diameter of the Kevlar string (strings that will be used to actuate the joints).</li>
		<li>Assemble all bone pieces together from base to tip using axles. Do this by placing all the components on a flat table as shown in Figure 4. To connect the base and middle piece, align the knuckles of the parts and insert the axle. Use the same technique to connect the middle and the tip piece together. Use an adhesive on each end of each axle to ensure the axle does not move laterally (Figure 5b).</li>
		<li>Cut plastic tubes to the following length. Cut four tubes the length of the base bone piece (L1 = 8 cm) and two tubes the length of the middle piece (L2 = 6 cm).</li>
		<li>Cut 4 pieces of Kevlar string, each 3 feet in length.</li>
		<li>Slide one string through an L1 tube and then an L2 tube. Slide another string through an L1 tube. Repeat the process with the remaining tubes and strings.</li>
		<li>Place the tubes on top of the bone structures and use a clear tape to hold them in position temporarily. Using an adhesive, stick the tubes onto the bone structure and then remove the tapes. 
		NOTE: There is no specific position in which the tubes have to be placed, the critical aspect is to just stick them on the surface of the structure. Use Figure 5c as a guideline.</li>
		<li>Thread the Kevlar string from L1 tube and L2 tube through the holes drilled onto the tip and middle pieces as described in step 3.1.2. Make a small but secure knot once the string is through the hole (Figure 5d).</li>
	</ol>
	</li>
	<li>Adding the skin of the flipper to create a final flipper.
	<ol>
		<li>Measure 200 mL of silicon and silicon medium in two different containers.</li>
		<li>Pour both these liquids into a steel bowl. Add paint thinner (not to exceed 10% of weight of the total mixture) to the mixture for easy pouring and mixing.</li>
		<li>Use a stand mixer to mix the mixture thoroughly for 3 - 4 min. Color can be added at this step to achieve the desired visual effects. If a stand mixer is not available, use a whisk to mix it, taking care to scrape the sides and bottom of the container.</li>
		<li>Insert a rod into the knuckles of the base part and align it with the knuckles of the flipper mold. When the pegs fit into the cavities of the mold, the bone structure is aligned perfectly in the flipper mold. While holding down on the two parts of mold, secure the parts by using a clamp for added compression (this step is critical so that the silicon mixture does not leak from the gap between the two parts).</li>
		<li>Once the mixture is mixed, carefully pour it in the mold till the topmost knuckles of the bone structure. Oozing of liquid from the bottom hole in the mold is a sign of the mixture getting uniformly distributed. At the onset of this, plug the hole to avoid further flow of the liquid. Leave the liquid to cure for four hours before removing the flipper robot from the mold (see Figure 6).</li>
	</ol>
	</li>
</ol>
4. Mounting
<ol>
	<li>To mount the silicon foreflipper on the water flume (Figure 7), create a mounting structure. A CAD representation of the finished assembly is shown. (Figure 8).
	<ol>
		<li>Design a plate with a carefully extruded cut using CAD software. Click on &#39;Sketch&#39; and draw a rectangle of dimensions 14 x 19 inches (the height does not matter as the laser cutter uses a .dwg file). Use a rectangular sheet of steel as the base to manufacture this plate. Upload a two-dimensional drawing from the CAD software on a computer attached to a steel laser cutter to attain the desired cuts. 
		NOTE: This plate houses the motor and the cut in it allows for the pulley system to work. The width of the plate is equal to the width of the water flume, thus making it easier to slide the plate over the flume. This type of placement helps in easy removal of the mounting assembly to replace parts or the foreflipper model.</li>
		<li>Fix the foreflipper and the pulley onto a shaft, which slides into a triangular truss. 
		NOTE: A three-pulley system is implemented to transfer the torque/power from the motor to the rod.</li>
		<li>Use bearings on either side to help the rod to rotate smoothly. To restrict the movement of rod in the lateral direction, place shaft collars on each end of the shaft.</li>
	</ol>
	</li>
	<li>Set the motion of the flipper by selecting the jogging function on the driver. Pressing the &#39;Up&#39; button rotates the flipper clockwise and the &#39;Down&#39; button rotates the flipper anticlockwise. The driver allows for change the revolutions per minute of the motor shaft according to the instructions in the manual20.</li>
	<li>Insert the right-angled dye port in the water and increase the pressure on the dye system. Adjust the speed of the dye to the freestream velocity of the water so the dye appears as a single smooth filament. Rotate the flipper so that the dye interacts and gets trapped with the resulting vortices generated.</li>
</ol>

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The authors have nothing to disclose.

The robotic flipper apparatus will allow us to understand the hydrodynamics of the swimming California sea lion. This includes the basic thrust producing stroke (the &#39;clap&#39;), as well as non-physical variations that animal studies cannot investigate. The robotic flipper has been designed for experimental versatility, thus, step 3&#8212;where the flipper itself is made&#8212;is critical in obtaining the desired results. While this apparatus is, clearly, just a model of the living system, in situ studies of...

While scientists have investigated the basic characteristics of sea lion swimming (energetics, cost of transport, drag coefficient, linear speed and acceleration1-3, we lack information about the fluid dynamics of the system. Without this knowledge, we limit potential high-speed, high-maneuverability engineering applications to body-caudal fin (BCF) locomotion models4. By characterizing a different swimming paradigm, we hope to expand our catalog of design tools, specifically those with the potential to enable quieter, stealthier forms of swimming. Thus, we study the fundamental mechanism of sea lion swimming through direct observation of the California sea lion and laboratory investigations using a robotic sea lion foreflipper5,6 .
To do this, we will employ a commonly used technique for exploring complex biological systems: a robotic platform7. Several locomotion studies&#8212;both of walking8,9 and swimming10&#8212;have been based on either complex11 or highly simplified12 mechanical models of animals. Typically, the robotic platforms retain the essence of the model system, while allowing researchers to explore large parameter spaces13-15. While not always characterizing the entire system, much is learned through these platforms that isolate a single component of a locomotive system. For example, the fundamental functioning of unsteady propulsors, like the back-and-forth sweeping of a caudal fin during carangiform swimming, has been intensely explored through experimental investigations of pitching and/or heaving panels12,16,17,18. In this case, we can isolate certain modes of this complex motion in ways that animal based studies cannot. Those fundamental aspects of propulsion can then be used in the design of vehicles which do not need the biological complexity evolution provides.
In this paper, we present a novel platform for exploring the &#39;clap&#39; phase of the sea lion thrust-producing stroke. Only a single foreflipper&#8212;the &#39;roboflipper&#39;&#8212;is included in the platform. Its geometry is derived exactly from biological scans of a California sea lion (Zalophus californianus) specimen. The roboflipper is actuated to replicate the motion of the animals&#39; derived from previous studies1. This robotic flipper will be used to investigate the hydrodynamic performance of the swimming sea lion and to explore a wider parameter space than animal studies, particularly those of large aquatic mammals, can yield.

<table><tbody><tr><td>Dragon Skin 20</td><td>Smooth-on</td><td></td><td></td></tr><tr><td>Dragon Skin 20 medium</td><td>Smooth-on</td><td></td><td></td></tr><tr><td>Object24</td><td>Stratasys</td><td></td><td>3D printer</td></tr><tr><td>Stand Mixer</td><td>Hamilton</td><td></td><td></td></tr><tr><td>PKS-PRO-E-10 System</td><td>Anaheim Automation</td><td>PKS-PRO-E-10-A-LP22</td><td>Controller and Servo Motor</td></tr><tr><td>Artec Eva</td><td>Artec 3D</td><td></td><td>3D light scanner with resolution of 0.1 mm</td></tr><tr><td>Artec Spider</td><td>Artec 3D</td><td></td><td>3D light scanner with resolution of 0.5 mm</td></tr><tr><td>Steel plate</td><td>Mcmaster</td><td></td><td></td></tr><tr><td>Carbon Tow</td><td>Fibreglast</td><td>2393-A</td><td></td></tr><tr><td>Hardened Precision 440C Stainless Steel Shaft</td><td>Mcmaster</td><td>6253K49</td><td></td></tr><tr><td>Tygon PVC Clear Tubing</td><td>Mcmaster</td><td>6546T23</td><td></td></tr><tr><td>Kevlar Thread</td><td>Mcmaster</td><td></td><td></td></tr></tbody></table>

a robotic platform to study the foreflipper of the california sea lion

The California sea lion (Zalophus californianus), is an agile and powerful swimmer. Unlike many successful swimmers (dolphins, tuna), they generate most of their thrust with their large foreflippers. This protocol describes a robotic platform designed to study the hydrodynamic performance of the swimming California sea lion (Zalophus californianus). The robot is a model of the animal&#39;s foreflipper that is actuated by motors to replicate the motion of its propulsive stroke (the &#39;clap&#39;). The kinematics of the sea lion&#39;s propulsive stroke are extracted from video data of unmarked, non-research sea lions at the Smithsonian Zoological Park (SNZ). Those data form the basis of the actuation motion of the robotic flipper presented here. The geometry of the robotic flipper is based a on high-resolution laser scan of a foreflipper of an adult female sea lion, scaled to about 60% of the full-scale flipper. The articulated model has three joints, mimicking the elbow, wrist and knuckle joint of the sea lion foreflipper. The robotic platform matches dynamics properties&#8212;Reynolds number and tip speed&#8212;of the animal when accelerating from rest. The robotic flipper can be used to determine the performance (forces and torques) and resulting flowfields.

The process described above yields a robotic model of a California sea lion foreflipper. The model can be used in two different ways. One is by actuating the flipper only at the root (Figure 6a). In this case, the driving motor sets the rotational rate of the first joint, but the resulting motion of the flipper is determined by the fluid-structure interaction between the flexible flipper and the surrounding water. Additionally, we can create robotic flipp...

Watch this Scientific Journal Video about A Robotic Platform to Study the Foreflipper of the California Sea Lion at JoVE.com

A Robotic Platform to Study the Foreflipper of the California Sea Lion

A robotic platform is described that will be used to study the hydrodynamic performance&#8212;forces and flowfields&#8212;of the swimming California sea lion. The robot is a model of the animal&#39;s foreflipper that is actuated by motors to replicate the motion of its propulsive stroke (the &#39;clap&#39;).

The California sea lion (Zalophus californianus), is an agile and powerful swimmer. Unlike many successful swimmers (dolphins, tuna), they generate most ...

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7.9K Views.  The George Washington University. A robotic platform is described that will be used to study the hydrodynamic performance—forces and flowfields—of the swimming California sea lion. The robot is a model of the animal's foreflipper that is actuated by motors to replicate the motion of its propulsive stroke (the 'clap').

Video: A Robotic Platform to Study the Foreflipper of the California Sea Lion

A Novel Method for Assessing Proximal and Distal Forelimb Function in the Rat: the Irvine, Beatties and Bresnahan (IBB) Forelimb Scale

Several experimental models of cervical spinal cord injury (SCI) have been developed recently to assess the consequences of damage to this level of the spinal cord (Pearse et al., 2005, Gensel et al., 2006, Anderson et al., 2009), as the majority of human SCI occur here (Young, 2010; www.sci-info-pages.com). Behavioral deficits include loss of forelimb function due to damage to the white matter affecting both descending motor and ascending sensory systems, and to the gray matter containing the segmental circuitry for processing sensory input and motor output for the forelimb. Additionally, a key priority for human patients with cervical SCI is restoration of hand/arm function (Anderson, 2004). Thus, outcome measures that assess both proximal and distal forelimb function are needed. Although there are several behavioral assays that are sensitive to different aspects of forelimb recovery in experimental models of cervical SCI (Girgis et al., 2007, Gensel et al., 2006, Ballerman et al., 2001, Metz and Whishaw, 2000, Bertelli and Mira, 1993, Montoya et al., 1991, Whishaw and Pellis, 1990), few techniques provide detailed information on the recovery of fine motor control and digit movement.
The current measurement technique, the Irvine, Beatties and Bresnahan forelimb scale (IBB), can detect recovery of both proximal and distal forelimb function including digit movements during a naturally occurring behavior that does not require extensive training or deprivation to enhance motivation. The IBB was generated by observing recovery after a unilateral C6 SCI, and involves video recording of animals eating two differently shaped cereals (spherical and doughnut) of a consistent size. These videos were then used to assess features of forelimb use, such as joint position, object support, digit movement and grasping technique.
The IBB, like other forelimb behavioral tasks, shows a consistent pattern of recovery that is sensitive to injury severity. Furthermore, the IBB scale could be used to assess recovery following other types of injury that impact normal forelimb function.

A Novel Method for Assessing Proximal and Distal Forelimb Function in the Rat: the Irvine, Beatties and Bresnahan IBB Forelimb Scale

Here we will describe a rodent behavioral assay that can detect recovery of both proximal and distal forelimb function including digit movements during a naturally occurring behavior that does not require extensive training or deprivation to enhance motivation.

Several experimental models of cervical spinal cord injury (SCI) have been developed recently to assess the consequences of damage to this level of the ...

Neuroscience

Flat-floored Air-lifted Platform: A New Method for Combining Behavior with Microscopy or Electrophysiology on Awake Freely Moving Rodents

It is widely acknowledged that the use of general anesthetics can undermine the relevance of electrophysiological or microscopical data obtained from a living animal&#8217;s brain. Moreover, the lengthy recovery from anesthesia limits the frequency of repeated recording/imaging episodes in longitudinal studies. Hence, new methods that would allow stable recordings from non-anesthetized behaving mice are expected to advance the fields of cellular and cognitive neurosciences. Existing solutions range from mere physical restraint to more sophisticated approaches, such as linear and spherical treadmills used in combination with computer-generated virtual reality. Here, a novel method is described where a head-fixed mouse can move around an air-lifted mobile homecage and explore its environment under stress-free conditions. This method allows researchers to perform behavioral tests (e.g., learning, habituation or novel object recognition) simultaneously with two-photon microscopic imaging and/or patch-clamp recordings, all combined in a single experiment. This video-article describes the use of the awake animal head fixation device (mobile homecage), demonstrates the procedures of animal habituation, and exemplifies a number of possible applications of the method.

This method creates a tangible, familiar environment for the mouse to navigate and explore during microscopic imaging or single-cell electrophysiological recordings, which require firm fixation of the animal&#8217;s head.

It is widely acknowledged that the use of general anesthetics can undermine the relevance of electrophysiological or microscopical data obtained from a ...

Behavior

Automated Interactive Video Playback for Studies of Animal Communication

Video playback is a widely-used technique for the controlled manipulation and presentation of visual signals in animal communication. In particular, parameter-based computer animation offers the opportunity to independently manipulate any number of behavioral, morphological, or spectral characteristics in the context of realistic, moving images of animals on screen. A major limitation of conventional playback, however, is that the visual stimulus lacks the ability to interact with the live animal. Borrowing from video-game technology, we have created an automated, interactive system for video playback that controls animations in response to real-time signals from a video tracking system. We demonstrated this method by conducting mate-choice trials on female swordtail fish, Xiphophorus birchmanni. Females were given a simultaneous choice between a courting male conspecific and a courting male heterospecific (X. malinche) on opposite sides of an aquarium. The virtual male stimulus was programmed to track the horizontal position of the female, as courting males do in the wild. Mate-choice trials on wild-caught X. birchmanni females were used to validate the prototype's ability to effectively generate a realistic visual stimulus.

Video playback is a widely used technique in animal behavior. We created and evaluated a program that applies rules-based, interactive playback of 3-D computer animations in response to real-time, automated data on subject behavior.

Video playback is a widely-used technique for the controlled manipulation and presentation of visual signals in animal communication. In particular, ...

Investigating Motor Skill Learning Processes with a Robotic Manipulandum

Skilled reaching tasks are commonly used in studies of motor skill learning and motor function under healthy and pathological conditions, but can be time-intensive and ambiguous to quantify beyond simple success rates. Here, we describe the training procedure for reach-and-pull tasks with ETH Pattus, a robotic platform for automated forelimb reaching training that records pulling and hand rotation movements in rats. Kinematic quantification of the performed pulling attempts reveals the presence of distinct temporal profiles of movement parameters such as pulling velocity, spatial variability of the pulling trajectory, deviation from midline, as well as pulling success. We show how minor adjustments in the training paradigm result in alterations in these parameters, revealing their relation to task difficulty, general motor function or skilled task execution. Combined with electrophysiological, pharmacological and optogenetic techniques, this paradigm can be used to explore the mechanisms underlying motor learning and memory formation, as well as loss and recovery of function (e.g. after stroke).

A paradigm is presented for training and analysis of an automated skilled reaching task in rats. Analysis of pulling attempts reveals distinct subprocesses of motor learning.

Skilled reaching tasks are commonly used in studies of motor skill learning and motor function under healthy and pathological conditions, but can be ...

The Knob Supination Task: A Semi-automated Method for Assessing Forelimb Function in Rats

Tasks that accurately measure dexterity in animal models are critical to understand hand function. Current rat behavioral tasks that measure dexterity largely use video analysis of reaching or food manipulation. While these tasks are easy to implement and are robust across disease models, they are subjective and laborious for the experimenter. Automating traditional tasks or creating new automated tasks can make the tasks more efficient, objective, and quantitative. Since rats are less dexterous than primates, central nervous system (CNS) injury produces more subtle deficits in dexterity, however, supination is highly affected in rodents and crucial to hand function in primates. Therefore, we designed a semi-automated task that measures forelimb supination in rats. Rats are trained to reach and grasp a knob-shaped manipulandum and turn the manipulandum in supination to receive a reward. Rats can acquire the skill within 20 &plusmn; 5 days. While the early part of training is highly supervised, much of the training is done without direct supervision. The task reliably and reproducibly captures subtle deficits after injury and shows functional recovery that accurately reflects clinical recovery curves. Analysis of data is performed by specialized software through a graphical user interface that is designed to be intuitive. We also give solutions to common problems encountered during training, and show that minor corrections to behavior early in training produce reliable acquisition of supination. Thus, the knob supination task provides efficient and quantitative evaluation of a critical movement for dexterity in rats.

This manuscript describes a semi-automated task that quantifies supination in rats. Rats reach, grasp, and supinate a spherical manipulandum. The rat is rewarded with a pellet if the turn angle exceeds a criterion set by the user. This task increases throughput, sensitivity to injury, and objectivity compared to traditional tasks.

Tasks that accurately measure dexterity in animal models are critical to understand hand function. Current rat behavioral tasks that measure dexterity ...

An In Vitro Preparation for Eliciting and Recording Feeding Motor Programs with Physiological Movements in Aplysia californica

Multifunctionality, the ability of one peripheral structure to generate multiple, distinct behaviors1, allows animals to rapidly adapt their behaviors to changing environments. The marine mollusk Aplysia californica provides a tractable system for the study of multifunctionality. During feeding, Aplysia generates several distinct types of behaviors using the same feeding apparatus, the buccal mass. The ganglia that control these behaviors contain a number of large, identified neurons that are accessible to electrophysiological study. The activity of these neurons has been described in motor programs that can be divided into two types, ingestive and egestive programs, based on the timing of neural activity that closes the food grasper relative to the neural activity that protracts or retracts the grasper2. However, in isolated ganglia, the muscle movements that would produce these behaviors are absent, making it harder to be certain whether the motor programs observed are correlates of real behaviors. In vivo, nerve and muscle recordings have been obtained corresponding to feeding programs2,3,4, but it is very difficult to directly record from individual neurons5. Additionally, in vivo, ingestive programs can be further divided into bites and swallows1,2, a distinction that is difficult to make in most previously described in vitro preparations.
The suspended buccal mass preparation (Figure 1) bridges the gap between isolated ganglia and intact animals. In this preparation, ingestive behaviors - including both biting and swallowing - and egestive behaviors (rejection) can be elicited, at the same time as individual neurons can be recorded from and stimulated using extracellular electrodes6. The feeding movements associated with these different behaviors can be recorded, quantified, and related directly to the motor programs. The motor programs in the suspended buccal mass preparation appear to be more similar to those observed in vivo than are motor programs elicited in isolated ganglia. Thus, the motor programs in this preparation can be more directly related to in vivo behavior; at the same time, individual neurons are more accessible to recording and stimulation than in intact animals. Additionally, as an intermediate step between isolated ganglia and intact animals, findings from the suspended buccal mass can aid in interpretation of data obtained in both more reduced and more intact settings. The suspended buccal mass preparation is a useful tool for characterizing the neural control of multifunctionality in Aplysia.

An In Vitro Preparation for Eliciting and Recording Feeding Motor Programs with Physiological Movements in Aplysia californica

We describe a technique to extracellularly record and stimulate from nerves, muscles, and individual identified neurons in vitro while eliciting and observing different types of feeding behaviors in the feeding apparatus of Aplysia.

Multifunctionality, the ability of one peripheral  structure to generate multiple, distinct behaviors1, allows animals  to rapidly adapt their behaviors ...

Recordings of Neural Circuit Activation in Freely Behaving Animals

The relationship between patterns of neural activity and corresponding behavioral expression is difficult to establish in unrestrained animals. Traditional non-invasive methods require at least partially restrained research subjects, and they only allow identification of large numbers of simultaneously activated neurons. On the other hand, small ensembles of neurons or individual neurons can only be measured using single-cell recordings obtained from largely reduced preparations. Since the expression of natural behavior is limited in restrained and dissected animals, the underlying neural mechanisms that control such behavior are difficult to identify. 
Here, I present a non-invasive physiological technique that allows measuring neural circuit activation in freely behaving animals. Using a pair of wire electrodes inside a water-filled chamber, the bath electrodes record neural and muscular field potentials generated by juvenile crayfish during natural or experimentally evoked escape responses. The primary escape responses of crayfish are mediated by three different types of tail-flips which move the animals away from the point of stimulation. Each type of tail-flip is controlled by its own neural circuit; the two fastest and most powerful escape responses require activation of different sets of large &#8220;command&#8221; neurons. In combination with behavioral observations, the bath electrode recordings allow unambiguous identification of these neurons and the associated neural circuits. Thus activity of neural circuitry underlying naturally occurring behavior can be measured in unrestrained animals and in different behavioral contexts.

Non-invasive measurements of neural activity patterns in freely behaving animals are obtained by combining neurophysiological recordings with high speed videography.

The relationship between patterns of neural activity and corresponding behavioral expression is difficult to establish in unrestrained animals. ...

Biology

Automated Rat Single-Pellet Reaching with 3-Dimensional Reconstruction of Paw and Digit Trajectories

Rodent skilled reaching is commonly used to study dexterous skills, but requires significant time and effort to implement the task and analyze the behavior. Several automated versions of skilled reaching have been developed recently. Here, we describe a version that automatically presents pellets to rats while recording high-definition video from multiple angles at high frame rates (300 fps). The paw and individual digits are tracked with DeepLabCut, a machine learning algorithm for markerless pose estimation. This system can also be synchronized with physiological recordings, or be used to trigger physiologic interventions (e.g., electrical or optical stimulation).

Rodent skilled reaching is commonly used to study dexterous skills, but requires significant time and effort to implement the task and analyze the behavior. We describe an automated version of skilled reaching with motion tracking and three-dimensional reconstruction of reach trajectories.

Rodent skilled reaching is commonly used to study dexterous skills, but requires significant time and effort to implement the task and analyze the ...

Cardiac Muscle-cell Based Actuator and Self-stabilizing Biorobot - PART 1

Biological machines often referred to as biorobots, are living cell- or tissue-based devices that are powered solely by the contractile activity of living components. Due to their inherent advantages, biorobots are gaining interest as alternatives to traditional fully artificial robots. Various studies have focused on harnessing the power of biological actuators, but only recently studies have quantitatively characterized the performance of biorobots and studied their geometry to enhance functionality and efficiency. Here, we demonstrate the development of a self-stabilizing swimming biorobot that can maintain its pitch, depth, and roll without external intervention. The design and fabrication of the PDMS scaffold for the biological actuator and biorobot followed by the functionalization with fibronectin is described in this first part. In the second part of this two-part article, we detail the incorporation of cardiomyocytes and characterize the biological actuator and biorobot function. Both incorporate a base and tail (cantilever) which produce fin-based propulsion. The tail is constructed with soft lithography techniques using PDMS and laser engraving. After incorporating the tail with the device base, it is functionalized with a cell adhesive protein and seeded confluently with cardiomyocytes. The base of the biological actuator consists of a solid PDMS block with a central glass bead (acts as a weight). The base of the biorobot consists of two composite PDMS materials, Ni-PDMS and microballoon-PDMS (MB-PDMS). The nickel powder (in Ni-PDMS) allows magnetic control of the biorobot during cells seeding and stability during locomotion. Microballoons (in MB-PDMS) decrease the density of MB-PDMS, and enable the biorobot to float and swim steadily. The use of these two materials with different mass densities, enabled precise control over the weight distribution to ensure a positive restoration force at any angle of the biorobot. This technique produces a magnetically controlled self-stabilizing swimming biorobot.

In this two-part study, a biological actuator was developed using highly flexible polydimethylsiloxane (PDMS) cantilevers and living muscle cells (cardiomyocytes), and characterized. The biological actuator was incorporated with a base made of modified PDMS materials to build a self-stabilizing, swimming biorobot.

Biological machines often referred to as biorobots, are living cell- or tissue-based devices that are powered solely by the contractile activity of living ...

Bioengineering

Flapping Soft Fin Deformation Modeling using Planar Laser-Induced Fluorescence Imaging

Propulsive mechanisms inspired by the fins of various fish species have been increasingly researched, given their potential for improved maneuvering and stealth capabilities in unmanned vehicle systems. Soft materials used in the membranes of these fin mechanisms have proven effective at increasing thrust and efficiency compared with more rigid structures, but it is essential to measure and model the deformations in these soft membranes accurately. This study presents a workflow for characterizing the time-dependent shape deformation of flexible underwater flapping fins using planar laser-induced fluorescence (PLIF). Pigmented polydimethylsiloxane fin membranes with varying stiffnesses (0.38 MPa and 0.82 MPa) are fabricated and mounted to an assembly for actuation in two degrees of freedom: pitch and roll. PLIF images are acquired across a range of spanwise planes, processed to obtain fin deformation profiles, and combined to reconstruct time-varying 3D deformed fin shapes. The data are then used to provide high-fidelity validation for fluid-structure interaction simulations and improve the understanding of the performance of these complex propulsion systems.

The present protocol involves the measurement and characterization of 3D shape deformation in underwater flapping fins built with polydimethylsiloxane (PDMS) materials. Accurate reconstruction of these deformations is essential for understanding the propulsive performance of compliant flapping fins.

Propulsive mechanisms inspired by the fins of various fish species have been increasingly researched, given their potential for improved maneuvering and ...

A Robotic Platform to Study the Foreflipper of the California Sea Lion

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Chapters in this video

A Novel Method for Assessing Proximal and Distal Forelimb Function in the Rat: the Irvine, Beatties and Bresnahan (IBB) Forelimb Scale

Flat-floored Air-lifted Platform: A New Method for Combining Behavior with Microscopy or Electrophysiology on Awake Freely Moving Rodents

Automated Interactive Video Playback for Studies of Animal Communication

Investigating Motor Skill Learning Processes with a Robotic Manipulandum

The Knob Supination Task: A Semi-automated Method for Assessing Forelimb Function in Rats

An In Vitro Preparation for Eliciting and Recording Feeding Motor Programs with Physiological Movements in Aplysia californica

Recordings of Neural Circuit Activation in Freely Behaving Animals

Automated Rat Single-Pellet Reaching with 3-Dimensional Reconstruction of Paw and Digit Trajectories

Cardiac Muscle-cell Based Actuator and Self-stabilizing Biorobot - PART 1

Flapping Soft Fin Deformation Modeling using Planar Laser-Induced Fluorescence Imaging