- Resources and references
- Blood vessel cannulation technique in other animals
- All blood sampling techniques in the mouse
Also referred to as peri-orbital, posterior-orbital and orbital venous sinus bleeding. Retro-orbital bleeding should typically be performed as a terminal procedure. It should only be used with recovery in rare circumstances with exceptional scientific justification (e.g. where a large blood volume is necessary or where peripheral veins are used for dosing) because of its potential impact on animal welfare. Adverse effects reported for this procedure are described in the summary below.
Where its use is unavoidable, retro-orbital bleeding should only be used under general anaesthesia. Because of the severity of the adverse effects that can occur with this technique, even in skilled hands, it is essential that it is conducted only by staff members competent (practiced) in the technique.
Blood is collected from the venous sinus. The mouse is restrained, the neck gently scruffed and the eye made to bulge. A capillary tube/pipette is inserted medially, laterally or dorsally. Blood is allowed to flow by capillary action into the capillary tube/pipette. The sample obtained is a mixture of venous blood and tissue fluid, and is not representative of venous blood.
Blood flow can be stopped by applying gentle finger pressure to the soft tissue. A finger should be placed over the closed eyelid for approximately 30 seconds. The mouse should be checked for post-operative peri-orbital lesions approximately 30 minutes after blood sampling and on at least one more occasion within two hours of the sampling.
Sequential sampling is not recommended as histological changes, abnormal clinical signs and evidence of discomfort have been reported. Ideally, no more than one bleed per eye should be taken - where a second retro-orbital bleed is required this should be performed as a terminal procedure. In rare circumstances where there is exceptional scientific justification sequential sampling may be used. In such instances only one orbit should be sampled from at any one sampling time. Adequate time should elapse between sampling to allow peri-orbital tissue repair to take place. An interval of two weeks between bleeds should allow damaged tissue to repair in most cases.
|Number of samples||It is recommended that only one sample be taken.|
|Sample volume||Up to 0.2 ml with recovery; Up to 0.5 ml non-recovery.|
|Equipment||A glass capillary tube or Pasteur pipette.|
|Staff resource||One person is required to take the blood sample.|
|Other||Careful monitoring for adverse effects is necessary both peri- and post-operatively.|
- A good practice guide to the administration of substances and removal of blood, including routes and volumes.
- Luzzi M et al. (2005) Collecting blood from rodents: a discussion by the Laboratory Animal Refinement and Enrichment Forum. Animal Technology and Welfare. 4(2) 99-102.
- Schnell MA, Hardy C, Hawley M, Propert KJ, Wilson JM (2002) Effect of blood collection technique in mice on clinical pathology parameters. Human Gene Therapy. 13(1) 155-161.
- Methods of blood collection in the mouse.
- Forbes N, Brayton C, Grindle S, Shepherd S, Tyler B, Guarnieri M (2010) Morbidity and mortality rates associated with serial bleeding from the superficial temporal vein in mice. Lab Animal Europe. 10(9), 14-22
- Removal of blood from laboratory animals and birds.