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Sampling of the retroorbital sinus is associated with tissue destruction & damage to periorbital structures, resulting in unnecessary pain and distress – especially when multiple samples are obtained from the same eye. The NIH guidelines express concern regarding timing and the use of alternate eyes:
- A minimum of 10 days should be allowed for tissue repair before repeat sampling from the same orbit. Otherwise the healing process may interfere with blood flow.
- Alternating orbits should not be attempted until the phlebotomist is proficient in obtaining samples from the orbit accessed most readily by the dominant hand i.e., a right handed individual should gain proficiency withdrawing samples from the right orbit before attempting to obtain samples from the left orbit.
As an alternative, the IACUC recommends that the facial/superficial temporal vein phlebotomy technique should be used in place of retro-orbital bleeding whenever possible. 150 µl of blood is readily obtainable using this method. Utilization of this method in place of retro-orbital bleeding is considered an element of the "three R's" of animal research - Refinement of techniques to reduce pain or distress.
Note: tail vein or artery phlebotomy is acceptable for smaller quantities of blood.
Below are links to further information and techniques. Please visit these on-line sites. If your lab personnel require additional training, please contact the IAS veterinary staff for assistance.
https://www.youtube.com/watch?v=niTVnEAHOko
http://www.medipoint.com/html/mouse_phlebotomy.html