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

ラットの採血技術の比較研究

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).

著者スポットライト:ラットにおける状況依存性の採血方法 

ラットモデルにおける採血のための修正尾静脈および陰茎静脈穿刺

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  

麻酔なしの尾静脈穿刺

はじめに、ラットをプラスチック製の拘束コーンに固定し、円錐の大きい方の端を尾の付け根の周りに囲みます。動物が快適で、手順全体を通して呼吸が制限されていないことを確認してください。尾を温水に約1分間浸して静脈を拡張し、ペーパータオルで尾を乾かします。拘束具で、尾を加熱パッドの上に横たえて動物を下に向けて配置します。サンプリングに適した尾静脈を選択するには、動物全体を右側または左側に回転させます。このゾーンでは血管がより表面的になるため、血管の穿刺には尾の末端3番目を使用します。穿刺部位で70%エタノールワイプで尾を拭き、加熱パッドの端に置き、尾の末端3分の1に角度を付けます。これにより、静脈が表面にもたらされ、サンプルを採取するためのスペースが増えます。採血の場合は、プランジャーをシリンジ内で数回前後に動かして引き出しをスムーズにしてから、プランジャーを引っ張ってシリンジ内に負圧をかけます。非利き手の人差し指と中指を使用して、テールを加熱パッドに平らに固定します。中指を近位に、人差し指を遠位に配置し、これら2本の指の間の穿刺部位を置きます。インスリン注射器を人差し指に対してスライドさせ、針の目が上を向いて静脈に挿入されるまでスライドさせます。これにより、針と尾の間に35度の角度が作成されます。針が静脈に入ると、血液が注射器に流れ込みます。シリンジのプランジャーを安定した速度でゆっくりと引き出して、目的の容量を収集します。シリンジを尾から取り外し、尾の穿刺側に血滴ができるようにします。非滅菌処置の場合、この血液を吸引してさらに数マイクロリットルの血液を収集します。穿刺部位に圧力を加えて出血を止め、新しいアルコールワイプでその領域を拭きます。ラットを円錐形から取り出した後、ケージに戻します。尾静脈穿刺法の成功率を麻酔下での陰茎静脈穿刺法の成功率と比較した。興味深いことに、麻酔下では、尾静脈は非常に信頼性が低くなり、成功率はわずか25%でした成功率は、意識のあるラットの尾静脈穿刺と麻酔下の陰茎静脈穿刺についてほぼ同様の結果を示しました。採血は麻酔なしの尾静脈穿刺法と麻酔下の陰茎静脈穿刺法で6分未満で行われたのに対し、麻酔下での尾静脈穿刺法は複数の失敗のために8分以上かかりました。

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 ...