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

Sammenlignende undersøgelse af blodopsamlingsteknikker hos rotter

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).

Forfatter Spotlight: Kontekstafhængige blodindsamlingsmetoder hos rotter 

Modificeret halevene og penisvenepunktur til blodprøvetagning i rottemodellen

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  

Halevenepunktur uden bedøvelse

Til at begynde med skal du sikre rotten i en plastikkegle, vedlægge den store ende af keglen omkring bunden af halen. Sørg for, at dyret er behageligt og vejrtrækning er ubegrænset under hele proceduren. Dyp halen i varmt vand i cirka et minut for at udvide venen, og tør halen med et papirhåndklæde.I fastholdelsen skal du placere dyret nedad med halen liggende på en varmepude. For at vælge den passende halevene til prøveudtagning skal du dreje hele dyret til højre eller venstre side. Brug den sidste tredjedel af halen til punktering af blodkar, da karrene bliver mere overfladiske i denne zone.Tør halen af med 70% ethanolservietter på punkteringsstedet, og placer den på kanten af varmepuden, hvilket skaber en vinkel i den sidste tredjedel af halen. Dette bringer venen til overfladen og skaber mere plads til at tage prøven. Ved blodprøvetagning skal du bevæge stemplet frem og tilbage i sprøjten flere gange for at udjævne udtagningen og derefter trække i stemplet for at skabe undertryk i sprøjten.Fastgør halen fladt på varmepuden ved hjælp af den ikke-dominerende pege- og langfinger. Placer langfingeren proximalt og pegefingeren distalt med punkteringsstedet mellem disse to fingre. Skub insulinsprøjten mod pegefingeren med kanyleøjet pegende opad, indtil det indsættes i venen.Dette skaber en vinkel på 35 grader mellem nålen og halen. Når nålen er kommet ind i venen, vil blodet strømme ind i sprøjten. Træk langsomt sprøjtens stempel op med en jævn hastighed for at opsamle det ønskede volumen.Fjern sprøjten fra halen og lad en bloddråbe dannes på punkteringssiden af halen. Aspirer dette blod til at indsamle et par flere mikroliter blod i tilfælde af en ikke-steril procedure. Tryk på punkteringsstedet for at stoppe blødningen, og tør området af med en ny alkoholserviet.Når du har fjernet rotten fra keglen, skal du returnere den til buret. Succesraterne med halevenepunkturmetoderne blev sammenlignet med succesraten for penisvenepunkturmetoden under anæstesi. Interessant nok blev halevenen under anæstesi meget upålidelig og havde en succesrate på kun 25%Succesraterne viste næsten lignende resultater for halevenepunktur hos bevidste rotter og penisvenepunktur under anæstesi.Blodprøvetagningen blev udført på mindre end seks minutter i halevenepunktur uden bedøvelse og penisvenepunkturmetoden under anæstesi, mens halevenepunkturmetoden under anæstesi tog mere end otte minutter på grund af flere fejl.

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 ...