Rat: Temporary cannula (non-surgical)

Technique

Temporary cannulation of the lateral tail vein should be considered when repeated blood samples are required over a short period of time (e.g. a few hours) as it avoids multiple needle entries and the associated damage to the tail vein. The technique may be combined with normal tail vein bleeding to accommodate all the blood samples required by a given protocol but reduce the number of needle entries and reduce the time the rat must spend in a warming cabinet (since warming may not be necesary for taking blood via the temporary cannula). It is suitable for use in all strains of rat, and the animals can be group-housed during the study period. 

Tail bleeding normally requires the rats to be warmed in order to dilate the blood vessel prior to taking the sample. This may be stressful and can cause dehydration due to salivation, in addition to increasing metabolic rate, which may affect the experimental data. Restraining the rat for long periods of time in a restraint tube may also cause it to become hot, which will be stressful to the rat and may affect the blood parameters. View this technique below.

Rat temporary cannula trimmed copyright NC3Rs from NC3Rs on Vimeo.

No surgery is required. An intravenous catheter is inserted into the vein by puncture of the skin and is taped in situ . A heparin flush is used (0.1 ml) after placement and between samples to prevent clotting. An access port is inserted into the exteriorised end of the cannula, which stops the blood from flowing, and the catheter is taped into place.

An aseptic technique should be used. A local anaesthetic cream (e.g. EMLA cream) can be applied to the site 30 minutes prior to insertion of the catheter. The tail may need to be washed with diluted Hibiscrub (1%) in order to see the blood vessel.

0.1 - 2.0 ml (normally 0.1 - 0.3 ml) can be taken per sample, and depending on the sample volume and consideration of the effects of repeated warming and restraint, no more than six samples in a two hour period or eight samples in a 24-hour period.

The lateral tail vein is usually accessed approximately one-third along the length of the tail from the tail tip. Cannulation should not be attempted at the base of the tail, as this could result in a perivascular clot and inflammation that significantly reduces blood flow to the distal portion of the vessel. If cannulation is unsuccessful, direct venepuncture may be used as the alternative. The number of attempts to take any blood sample should be minimised (no more than three needle sticks in any one attempt).

A warming cabinet is used prior to the cannulation (39oC for five to 15 minutes). Subsequent warming prior to sampling may not be required so long as blood is 'free flowing'. If necessary, the rat can be warmed for a short period (five minutes) as required. The rat should be carefully monitored, including checking for signs of hyperthermia and dehydration. The time the rat is in the warming cabinet should be recorded and the cabinet should be calibrated regularly to avoid hyperthermia; digital displays should not be relied upon. It is important to ensure the temperature in the cabinet is uniform and that there are no 'hot spots'. Alternatively, a warm bath at a maximum of 40oC can be used to warm just the tail of the rat. The temperature of the bath should be monitored otherwise the tail can be scalded.

Rats need to be restrained which can cause stress and therefore the duration of restraint should be minimised. Restraint can either be manual (e.g. wrapping the rat in a towel) or using a restraint tube. Anecdotal evidence suggests that holding the rat is less stressful than using a restraint tube. Where a restraint tube is used, it should be appropriate to the size of the rat in order to avoid damage to the tail, testes, limbs and back. All forms of restraining equipment should be frequently washed to prevent pheromonally-induced stress or cross-infection.

Summary

Number of samples Ideally, no more than six samples should be taken in a two hour period, or eight samples in a 24-hour period, depending on sample volume and consideration of the effects of repeated warming and restraint. 
Sample volume 0.1 - 2.0 ml (normally 0.1 - 0.3 ml)
Equipment 22G 0.90 mm i.v. catheter
Staff resource One person is required to take the blood sample if a tube restrainer is used. Two people would be required if one were to restrain the rat manually. For large batches of animals, two people are required: one person to take the blood sample and one to operate the warming cabinet.
Adverse effects
  • Bruising
  • Infection <1%
  • Haemorrhage <1%
Other Rats are warmed to dilate the blood vessel and care should be taken to avoid hyperthermia and dehydration. The time the animal is exposed to warming and restraint should be kept to a minimum.

Resources and references

  • A good practice guide to the administration of substances and removal of blood, including routes and volumes. 
  • Staszyk C, Bohnet W, Gasse H, Hackbarth H (2003), Blood vessels of the rat tail: a histological re-examination with respect to blood vessel puncture methods. Laboratory Animals. 37(2),  pp 121-125
  • McClure DE (1999), Clinical pathology and sample collection in the laboratory rodent. The Veterinary Clinics of North America: Exotic Animal Practice. 2(3),  pp 565-590, vi
  • Ness RD (1999), Clinical pathology and sample collection of exotic small animals. The Veterinary Clinics of North America: Exotic Animal Practice. 2(3),  pp 591-620
  • Lucas RL, Lentz KD, Hale AS (2004), Collection and preparation of blood products. Clinical Techniques in Small Animal Practice. 19(2),  pp 55-62
  • Removal of blood from laboratory animals and birds.

Temporary cannulation in other animals

This technique is only appropriate for use in the rat.

All blood sampling techniques in the rat

Click here for information on blood vessel cannulation in the ratClick here for information on tail vein blood sampling in the ratClick here for information on jugular vein blood sampling in the ratClick here for information on saphenous vein blood sampling in the ratClick here for information on retro-orbital blood sampling techniques in the ratClick here for information on abdominal/thoracic blood vessel blood sampling in the ratClick here for information on cardiac puncture blood sampling in the ratClick here for information on schedule 1 stunning followed by decapitation for blood sampling in the ratClick here for information on decapitation blood sampling techniques in the rat