Name: Drawing Blood from the Penile Vein
Uploaded: 2023-06-30T14:30:00
Description: Watch this Scientific Journal Video about Drawing Blood from the Penile Vein at JoVE.com

drawing blood from the penile vein

Studio comparativo delle tecniche di raccolta del sangue nei ratti

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).

Autore in primo piano: Metodi di raccolta del sangue dipendenti dal contesto nei ratti 

Puntura modificata della vena della coda e della vena del pene per il prelievo di sangue nel modello di ratto

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 Drawing Blood from the Penile Vein at JoVE.com

Drawing Blood from the Penile Vein  

Prelievo di sangue dalla vena del pene

Spostare lo stantuffo avanti e indietro nella siringa più volte per attenuare il prelievo e quindi tirare lo stantuffo per creare una pressione negativa nella siringa. Ritrarre il prepuzio del ratto anestetizzato dalla fine del pene e tenere le ghiandole tra l'indice e il pollice tirando delicatamente. La vena dorsale del pene apparirà come un cordone blu superficiale.Con la cruna dell'ago rivolta verso l'alto, inserire la siringa da insulina nella vena con un angolo di 35 gradi. Una volta che l'ago è entrato nella vena, il sangue fluirà nella siringa. Estrarre lentamente lo stantuffo della siringa a una velocità lenta e costante per raccogliere il volume desiderato.Rimuovere la siringa. Una goccia di sangue si formerà sul sito di puntura. Aspirare la goccia per raccogliere qualche microlitro in più di sangue in caso di procedura non sterile.Applicare una leggera pressione sul sito di puntura per fermare l'emorragia e pulire l'area con una nuova salvietta imbevuta di alcool. Rimetti il pene nella sua posizione neutra. Dopo aver spento l'isoflurano, monitorare il ratto fino al completo recupero e riportarlo nella sua gabbia.Il volume di sangue raccolto nel metodo di puntura della vena del pene dorsale in anestesia è stato confrontato con il sangue raccolto nel metodo di puntura della vena della coda con e senza anestesia. Il risultato mostra che il metodo di puntura della vena del pene in anestesia e il metodo di puntura della vena della coda senza anestesia hanno prodotto quantità comparabili di sangue.

Research

JoVE Journal

Medicine

Drawing Blood from the Penile Vein

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 ...