subclavian vein puncture blood collection and observation of secondary outcome measures in rats

锁骨下静脉穿刺法对清醒大鼠重复采血

Rats are one of the most common experimental animals, and there are many ways to obtain blood samples. For experiments involving a single blood collection at the concluding stage, a sufficient amount of blood can be obtained through cardiac puncture or abdominal aorta blood collection1. However, some studies require repeated blood collection from rats for routine blood or biochemical analysis, especially in pharmacokinetics and toxicology studies, where repeated blood collection is required to determine the absorption, distribution, and metabolism of drugs2.
Currently, although tail vein blood collection is the most common method for blood sampling from rats, despite not requiring anesthesia, this method can be challenging for repeated collections, and the volume of blood collected is relatively small3,4. Additionally, although blood can be collected from the saphenous and penile veins, the amount of blood obtained is limited, and anesthesia is required1,5. Furthermore, blood samples collected from the submandibular venous plexus, as well as sublingual, jugular, and subclavian veins provide higher-quality samples but typically require anesthesia or the assistance of multiple individuals1,6,7,8,9. Finally, retro-orbital sinus/canal blood collection not only necessitates anesthesia but may also potentially cause injury and stress to the rats9.
The quality of blood samples typically obtained from major veins is generally of the highest standard1. At present, some studies have found that continuous microsampling through the jugular vein is a very suitable method for toxicological research in rats although this method usually requires jugular vein catheterization10,11,12. Therefore, it is worth exploring how to obtain high-quality blood samples in accordance with the 3R principle of animal research without surgical intervention. The objective of this study was to present a method for efficiently extracting blood from the subclavian vein in rats. This technique enables the swift collection of satisfactory samples through a single-person procedure without the need for anesthesia.

作者聚焦：在非麻醉大鼠中开发一种高效的血液采集方法 

有意识大鼠的锁骨下静脉血液采样

The scope of our research is centered on devising infection methodologies for collecting blood samples from rats. Our aim is to ensure rapid and set procedures that align with our principle, utilizing minimal manpower and resources. One of the current experimental challenges is to effectively immobilize the rat in a manner that simultaneously maintains the subclavian vein enclosed proximately to the skin surface for optimal blood collection.The advantages of our protocol is that high quality blood samples can be obtained quickly by a person without anesthesizing the rats.

Watch this Scientific Journal Video about Subclavian Vein Puncture Blood Collection and Observation of Secondary Outcome Measures in Rats at JoVE.com

Subclavian Vein Puncture Blood Collection and Observation of Secondary Outcome Measures in Rats  

锁骨下静脉穿刺采血和大鼠次要结果测量的观察

在采血当天，在上午8：00将大鼠放入笼子中。称量食物和水的重量，然后将它们放入笼子中。根据以下标准定义成功的血液采样。让盲法研究人员每天上午 8：00 记录大鼠的体重、食物消耗量和饮水量。接下来，轻轻抚摸老鼠的背部皮肤，反复捏住它的脖子，帮助老鼠放松。使用非惯用手的拇指和食指，握住并提起大鼠的颈部皮肤。用剩下的三个手指和非惯用手的手掌，固定大鼠的背部皮肤，并在惯用手的帮助下，固定其前肢。用非惯用手的食指，轻轻向下推大鼠的头部皮肤。用其他手指沿手掌向外旋转肩关节，同时惯用手完全伸展肩关节。用非惯用手牢牢抓住大鼠，使其头部和身体对齐成一条直线。用惯用手定位锁骨并确认注射部位。用酒精对部位进行消毒，然后用肝素钠冲洗注射器。将注射器与大鼠的身体平行，惯用手，针尖朝上，注射器秤朝向实验者。将针插入锁骨切迹下方 0.5 厘米处。轻轻抽出注射器以产生负压，以恒定速度收集 0.1 至 1 毫升血液。用棉签止血，然后将大鼠放回笼子里。A 组和 B 组的平均采血时间在 96 小时内相似。A组采血成功率为93.8%，B组采血成功率为95.8%，差异无统计学意义。与第四天相比，A组第二天的平均采血时间较短。A组成功采血的次数平均为1.21次，B组为1.17次。两组在四天内都显示出稳定的体重增加。同性大鼠的水和食物摄入量保持相对稳定。

subclavian-vein-puncture-blood-collection-observation-secondary

Research

JoVE Journal

Medicine

Subclavian Vein Puncture Blood Collection and Observation of Secondary Outcome Measures in Rats

224 次观看.  Lanzhou University Second Hospital. 在采血当天，在上午8：00将大鼠放入笼子中。称量食物和水的重量，然后将它们放入笼子中。根据以下标准定义成功的血液采样。让盲法研究人员每天上午 8：00 记录大鼠的体重、食物消耗量和饮水量。接下来，轻轻抚摸老鼠的背部皮肤，反复捏住它的脖子，帮助老鼠放松。使用非惯用手的拇指和食指，握住并提起大鼠的颈部皮肤。用剩下的三个手指和非惯用手的手掌?...

视频: Subclavian Vein Puncture Blood Collection and Observation of Secondary Outcome Measures in Rats  

Subclavian Vein Blood Sampling in Conscious Rats

There are several established methods for obtaining repeated blood samples from rats, with the most commonly employed methods being lateral tail vein sampling without anesthesia and jugular vein sampling with anesthesia. However, most of these methods require assistance and anesthetic equipment and sometimes pose difficulties in terms of blood collection or the poor quality of blood samples. In addition, these methods of blood collection consume significant time and human resources when repeated blood sampling is required for a large number of rats. This study presents a technique for repetitive blood sampling in non-anesthetized rats by a single proficient individual. Highly satisfactory blood samples can be obtained by puncturing the subclavian vein. The method demonstrated an impressive overall success rate of 95%, with a median time of merely 2 min from rat restraint to the completion of blood collection. Furthermore, performing consecutive blood collections within the designated range does not inflict any harm on the rats. This method is worth promoting for blood collection, especially in large-scale pharmacokinetic studies.

Here we present a combination of effective rat restriction and subclavian vein puncture methods that enable rapid, safe, and repeated blood collection in rats without anesthesia.

There are several established methods for obtaining repeated blood samples from rats, with the most commonly employed methods being lateral tail vein ...

科研

医学

On the day of the blood sampling, place the rat in the cage at 8:00 AM.Weigh the food and water and place them in the cage. Define successful blood sampling based on the following criteria. Have a blinded researcher record the rat's weight, food consumption, and water intake daily at 8:00 AM.Next, gently stroke the rat's back skin and pinch its neck repeatedly to help the rat relax.Using the non-dominant hand's thumb and index finger, grip and lift the rat's neck skin. Using the remaining three fingers and the palm of the non-dominant hand, secure the rat's back skin, and with the help of the dominant hand, immobilize its front limbs. Using the non-dominant hand's index finger, gently push down on the rat's head skin.Rotate the shoulder joint outwardly with the other fingers along the palm while the dominant hand fully extends the shoulder joint. Grip the rat firmly with the non-dominant hand to align its head and body in a straight line. Locate the clavicle with the dominant hand and confirm the injection site.Disinfect the site with alcohol, then flush the syringe with sodium heparin. Hold the syringe parallel to the rat's body and the dominant hand with the needle tip facing upwards and the syringe scale toward the experimenter. Insert the needle 0.5 centimeters below the clavicle notch.Gently withdraw the syringe to create negative pressure, collecting 0.1 to 1 milliliters of blood at a constant speed. Apply a cotton swab for hemostasis and return the rat to its cage. Average blood collection times for groups A and B were similar over 96 hours.Blood collection success rates were 93.8%for group A and 95.8%for group B, with no significant difference. Group A had a shorter average blood collection time on the second day compared to the fourth day. The number of attempts for successful blood collection averaged 1.21 for group A and 1.17 for group B.More attempts and longer puncture times indicated higher failure rates.Both groups showed steady weight gain over four days. Water and food intake remained relatively constant among rats of the same sex.