Thanks to the Department of Neurosurgery of the Second Affiliated Hospital of Army Medical University for site support.&#160;Funding: This work was supported by the Natural Science Foundation of China (82104696).

The experiments in this study comply with the &#34;3Rs&#34; principle of animal ethics and have been approved by the Laboratory Animal Welfare and Ethnic Committee of the Army Medical University (AMUWEC20234543).
1. Preparation of rodent body fixation device
<ol>
	<li>Design a small animal holder based on the Chinese utility model patent9.</li>
	<li>Produce the device through 3D printing technology.</li>
	<li>Prepare a 2 m long rope and divide it into...

Using a 3D Printed Holder for Bulk Electroacupuncture in Small Animals

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The authors declared that no competing conflicts of interest exist.

Electroacupuncture (EA) is a form of acupuncture that involves the use of electrical stimulation on acupuncture needles11. This technique involves the use of micro-pulse currents of specific intensity and frequency to stimulate acupuncture points and achieve enhanced therapeutic effects3. However, the mechanism of EA for healing is limited and requires extensive basic research to prove1,2,<sup class="xref...

Electroacupuncture (EA) therapy is a unique method in which acupuncture needles are inserted into the scalp and connected to an electro-machine to stimulate specific points1. Unlike manual stimulation, EA allows better control of stimulation by stabilizing specific frequencies and waveforms to achieve optimal therapeutic effects2. According to a survey, 81.2% of medical institutions in China use EA or manual acupuncture to treat cerebral palsy, neuralgia, facial palsy, and other conditions. Despite its popularity, the specific mechanism of efficacy of EA is still unknown, which has hindered its promotion and application in the rehabilitation treatment of neurological diseases1,2,3. Further research is needed to fully understand the mechanism of EA&#39;s effectiveness and to promote its use in the rehabilitation treatment of neurological diseases.
As the influence of acupuncture expands worldwide, many studies have already investigated the mechanisms of EA performed on rodent models, such as rat or mouse models1,2,3. Several issues are often encountered in EA&#39;s rodent experiments. The first is how to immobilize rodents without anesthesia, as acupuncture performed on awake subjects is more reflective of clinical practice. In addition, some experiments require animals to be awake to observe treatment effects2,3. Another challenge is accurately locating the acupoints in mice or rats that correspond to those in humans. The precise localization of acupoints in experimental rodents is currently being studied by many scholars4,5. In this protocol, MS6 and ST25 were selected, which have been clearly defined in rodents by the transformation of human anatomical localization. MS6 is commonly used for the treatment of some brain diseases, such as Parkinson&#39;s disease6. ST25 is usually used to treat gastrointestinal problems, such as diarrhea7. These two acupoints were chosen primarily to demonstrate how rodents can be effectively immobilized in both supine and prone positions. Moreover, these acupoints have been extensively studied and offer significant insights for research purposes6,7.
The previous method of immobilizing a single rat for experiments is not only time-consuming but also difficult to handle by a single person8. Additionally, due to the non-cooperation of animals, the success rate is relatively low in practice. Therefore, there is a critical need to create an easily established animal model with stable characteristics to improve experiment efficiency. In this article, a 3D printed holder for small animals was introduced that could easily immobilize multiple rodents, leading to motor restriction. The aim of this paper is to administer EA treatments to a batch of young rats or mice, focusing on the strategies for bulk restriction of mice, identification, and stimulation of MS6 and ST25 acupoints.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>3D printing batch mice fixator</td>
			<td>MESH INVENT</td>
			<td>Custom made</td>
			<td>The fixator produced by 3D printer. The printing method is called fused deposition modeling (FDM), also known as fused filament fabrication (FFF).&#160;&#160;</td>
		</tr>
		<tr>
			<td>Electroacupuncture instrument</td>
			<td>Hwato, Suzhou Medical Appliance Factory</td>
			<td>SDZ-III</td>
			<td>www.Hwato-med.com</td>
		</tr>
		<tr>
			<td>Disposable sterile acupuncture needle</td>
			<td>Suzhou Medical Appliance Factory</td>
			<td>N/A</td>
			<td>0.25 mm x 13 mm</td>
		</tr>
	</tbody>
</table>

bulk electroacupuncture operation for mice or young rats

Electroacupuncture (EA) is widely used to treat various health conditions. However, the underlying mechanism of EA treatment remains unclear, hindering its promotion. The mechanistic study requires mouse or rat models to address this issue. However, these animals are not obedient to the experimental process, which is time-consuming. To solve these problems, we designed a 3D-printed small animal body bulk fixator to improve the efficiency of EA's animal experiments. This video shows in detail how to use the fixator to perform bulk EA on mice or young rats. For the selection of acupoints, the anterior oblique line of vertex temporal (MS6 head) and Tianshu point (ST25 belly) were chosen to verify the effect of the fixation device with prone positioning and supine positioning. Using the 3D-printed small animal holder allows multiple rodents to be immobilized and treated simultaneously, reducing the time and resources required for the experiment. This technique could be applied to other animal models by 3D printing different sizes and could potentially be used for various fixing conditions. The device is beneficial for the promotion of experimental scientific research in EA.

Author Spotlight: Using a 3D-Printed Holder for Simultaneous Multiple Electroacupuncture Operation

Bulk Electroacupuncture Operation for Mice or Young Rats

In this video, we introduce an efficient way of using electroacupuncture on multiple awake mice or rats. In vesical research, animal is urinating, but they are not cooperative, and acupuncture only is time consume. Another issue is about the anesthesia, which is not clinical applicable of acupuncture.To address these issues, we demonstrate efficient method of performing electroacupuncture on multiple awake mice or rats by using 3D printed holder. Our protocol demonstrates a more efficiency and safer way to hold multiple animals simultaneously, which unlike other method that imply immobilization, this method not only saves time, but also improve experimental success rates. Additionally, device is designed to be easy to handle, which could be operated by one person.The ProTech and device are beneficial for the promotion of experimental scientific research in electroacupuncture.

The floor plan of the animal body fixator we designed is shown in Figure 1. In addition, a 3D graphic model of this fixator was submitted to provide a comprehensive view of the design (Supplementary File 1). This is a device that allows 3 rodents to be immobilized and perform EA simultaneously. Rats and mice were restricted to the fixator in the awake state, and both prone and supine positions could be firmly fixed without causing injury to the rodents (...

Watch this Scientific Journal Video about Bulk Electroacupuncture Operation for Mice or Young Rats at JoVE.com

Here, we present a protocol for efficient and rapid electroacupuncture (EA) in mice or young rats using a three-dimensional (3D) printed holder. This technique allows simultaneous operation on multiple animals, saving time and increasing experimental efficiency.

Electroacupuncture (EA) is widely used to treat various health conditions. However, the underlying mechanism of EA treatment remains unclear, hindering ...

bulk-electroacupuncture-operation-for-mice-or-young-rats

Research

JoVE Journal

Biology

Electroacupuncture of Multiple Rodents in a 3D Printed Fixator

To begin, divide a two-meter long rope into six equal segments. Use the spring-loaded buckle mechanism to fasten the ends of the ropes, allowing for easy adjustment of loop sizes. Thread each of the six ropes through the designated holes in the fixator.Next position the rodent's head within the U-shaped hole in the fixator. Carefully fasten the loops around the rodent's neck and gently guide its body through two of the prepared rope loops. Then fasten the loops around the rodent's abdomen, ensuring it is tight enough without causing respiratory distress.Similarly immobilize the other two rodents. Find the acupoints by anatomical location and confirm the insertion depth at the acupoint. Slowly twist and insert the disposable sterile needles bilaterally and subcutaneously into MS6 or the ST25.Next, connect one side of the connection cable to the electroacupuncture instrument. Connect the other side of the connection cable with the positive and negative clips to the acupuncture needle shanks on the left and right MS6. Turn on the electroacupuncture instrument.Rotate the function or choose switch button to set the frequency and function mode. Set the frequency to 10 Hertz and choose a continuous wave mode. Rotate the Adjust Intensity button to set the intensity at one to two milliamperes lasting for five minutes.The 3D-printed fixator allowed for the simultaneous firm immobilization of three mice in both supine and prone positions without injury. The MS6 and ST25 acupoints were successfully located and needled in rodents. The average time for immobilization of three rats in both positions was between 72 to 77 seconds.

421 Views.  Army Medical University.  Here, we present a protocol for efficient and rapid electroacupuncture (EA) in mice or young rats using a three-dimensional (3D) printed holder. This technique allows simultaneous operation on multiple animals, saving time and increasing experimental efficiency.

Using a 3D Printed Holder for Bulk Electroacupuncture in Small Animals

Summary