Name: Tail Vein Puncture Without Anesthesia
Uploaded: 2023-06-30T14:30:00
Description: Watch this Scientific Journal Video about Tail Vein Puncture Without Anesthesia at JoVE.com

tail vein puncture without anesthesia

大鼠采血技术的比较研究

Blood sampling is necessary for most animal studies, both in vivo and in vitro. In rats, as the frequency and amount of blood sampling can be significant, it is helpful to have different alternatives for collection. Various methods have been described in previous studies.
The most commonly used techniques are tail vein puncture and saphenous vein blood draw. Tail vein sampling is suitable for all rat strains. With proper training, the procedure is simple to perform and causes minimal distress to the animal1. Similarly, the saphenous vein blood draw, provided it is done properly, is also a quick and simple collection method. Neither method requires anesthesia, and both allow for repeated draws of small amounts of blood. However, the saphenous vein puncture usually yields a lower blood volume1 and requires the presence of two people to leave one hindlimb exposed for puncture2.
If large amounts of blood need to be collected from a single animal, cardiac puncture or puncture of the vena cava may be used (up to 10 mL of blood can be drawn from a 150 g rat with cardiac puncture2). These techniques require anesthesia and are terminal procedures. The animal must be euthanized after any of these two techniques2. The jugular stick is an alternative that can be used if large amounts of blood need to be collected in a study that has not yet reached its endpoint. However, this technique also requires significant technical skills to avoid harm to the animal; hence, its use should be limited3.
Other techniques, such as the subclavian vein blood draw, do not need the use of anesthetics before blood collection and allow for repeated sampling of small volumes of blood. However, restrained handling and appropriate needle incision are required for this technique. An improper operation may result in animal pain or even mortality, and the training for this method may be fastidious4. 
Other anecdotal procedures include the orbital puncture and the sublingual vein puncture, both of which require an anesthetic, and neither are recommended nor widely used. Though previous studies have shown faster blood collection by orbital puncture than by tail vein puncture, it was found that orbital puncture under diethyl-ether anesthesia was less well tolerated than the latter method (based on the animals' excitation scores and urine production)5. Moreover, this method is highly influenced by the skill of the person who performs the procedure and is mainly performed by experienced veterinarians. Comparably, the sublingual vein puncture is less distressing and is recommended for repeated blood sampling6. However, this technique presents severe adverse effects such as reduced food and water intake, which can lead to the animal's death7. 
This study describes two methods used in our laboratory for repeated blood sampling. Tail vein puncture can be performed on a conscious animal, and the tissue damage and adverse effects are minimal. The modification of this technique in this study includes stabilizing the tail with the index and middle finger, which allows a single operator to perform the blood collection. The dorsal penile vein puncture has already been described for simple intravenous injections. This technique is performed under anesthesia and allows for a reliable blood source in case of difficulties with other methods (e.g., during the immediate postoperative period, with a small animal, when performing perioperative blood draw under anesthesia). Similar to tail vein sampling, the injury at the puncture site will have a minor overall effect on the animal compared to the techniques mentioned above8. The aim of this methods paper is to offer inexperienced researchers simple and reliable blood sampling alternatives according to the context (e.g., for procedures done under anesthesia, for studies including male rats, for animals with thin tail veins).

作者聚焦：大鼠的上下文依赖性采血方法 

改良尾静脉和静脉穿刺用于大鼠模型中的血液采样

The issue of repeated blood collection methods needs to be addressed since the majority of studies on rats require them. It's important to take into account that a sampling method can be very efficient in a certain context, but become unsuitable for another study. This protocol aims to offer simple and reliable blood sampling alternatives to researchers.Both methods can be done by a single operator and require no extensive prior training, which allows for standardized procedure. When an animal is under anesthesia, the effect of isoflurane can lead to vessel spasm and make the tail vein puncture unsuitable for blood collection. Our protocol offers an alternative in this situation, the penile vein puncture, which presents a better success rate and yields significantly more amounts of blood in less time.

Watch this Scientific Journal Video about Tail Vein Puncture Without Anesthesia at JoVE.com

Tail Vein Puncture Without Anesthesia  

无麻醉的尾静脉穿刺

首先，将大鼠固定在塑料约束锥中，将锥体的大端包围在尾巴的底部周围。确保动物在整个过程中舒适且呼吸不受限制。将尾巴浸入温水中约一分钟以扩张静脉，然后用纸巾擦干尾巴。在约束器中，将动物面朝下，尾巴放在加热垫上。要选择合适的尾静脉进行采样，请将整个动物向右侧或左侧旋转。使用尾巴的末端三分之一进行血管穿刺，因为血管在这个区域变得更加浅表。在穿刺部位用70%乙醇湿巾擦拭尾巴，并将其放在加热垫的边缘，在尾巴的末端三分之一处形成一个角度。这将静脉带到表面，并为取样创造更多空间。对于血液采样，将柱塞在注射器中来回移动几次以平滑退出，然后拉动柱塞以在注射器中产生负压。用非惯用食指和中指将尾巴平放在加热垫上。将中指放在近端，食指远端放置，穿刺部位放在这两个手指之间。将胰岛素注射器滑过食指，针眼朝上，直到插入静脉。这会在针和尾部之间产生 35 度角。一旦针头进入静脉，血液就会流入注射器。以稳定的速率缓慢抽出注射器的柱塞以收集所需的体积。从尾部取出注射器，让血滴在尾巴的穿刺侧形成。在非无菌程序的情况下，吸出这些血液以收集更多微升的血液。对穿刺部位施加压力以止血，并用新的酒精湿巾擦拭该区域。将大鼠从锥体中取出后，将其放回笼子。将麻醉下尾静脉穿刺法的成功率与静脉穿刺法的成功率进行比较。有趣的是，在麻醉下，尾静脉变得非常不可靠，成功率仅为25%，清醒大鼠尾静脉穿刺和麻醉下静脉穿刺的成功率几乎相似。无麻醉尾静脉穿刺法和麻醉静脉穿刺法不到6分钟，麻醉下尾静脉穿刺法因多次失败，抽血8分钟以上。

Research

JoVE Journal

Medicine

Tail Vein Puncture Without Anesthesia

Modified Tail Vein and Penile Vein Puncture for Blood Sampling in the Rat Model

Blood samples are required in most experimental animal designs to assess various hematological parameters. This paper presents two procedures for blood ...