Retro orbital bleeding protocol

Retro orbital bleeding protocol

Due to the nature and anatomy of rat tissues, retro-orbital bleeding may NEVER be performed in an unanesthetized rat. 4. It is best to alternate eyes in successive bleeds. Do not bleed from a damaged (ie. abscessed or ruptured) eye. In the event that one eye is damaged, the second eye can be bled. • Retro-orbital bleeding can be conducted in awake mice if a topical ophthalmic anesthetic is applied prior to the procedure. Alternatively, systemic anesthesia should be considered if compatible with the experimental design. • Due to pain and distress issues, retro-orbital sampling in the rat is best conducted under general anesthesia. Retroorbital bleeding can not be performed in awake mice unless documented and referenced scientific justification is provided in the submitted IACUC protocol. Rats must be fully anesthetized due to the presence of a plexus rather than a sinus which can lead to greater orbital tissue damage than in the mouse. institutions recommend the use of retro-orbital bleeding only for specific applications and encourage investigators to use other techniques, whereas other institutions place no restrictions on retro-orbital bleeding performed by trained investigators. It is the responsibility of both the investigator and IC ACUC

Collection from the retro-orbital venous plexus can cause several complications, and should be regarded as a technique requiring special justification and a high level of skill and competence. It is not recommended for routine use, but where it is used it must first have been shown that all other methods are inappropriate. Collection from the retro-orbital venous plexus can cause several complications, and should be regarded as a technique requiring special justification and a high level of skill and competence. It is not recommended for routine use, but where it is used it must first have been shown that all other methods are inappropriate. 6. For retro-orbital blood sampling: if the animal reaches a lighter plane of anesthesia, evidenced by increased respiratory rate, whisker twitch, or purposeful movement, stop the procedure and apply pressure to the eye to control any bleeding. Transfer the animal back to the bell jar, until the animal again reaches a deep plane of anesthesia. institutions recommend the use of retro-orbital bleeding only for specific applications and encourage investigators to use other techniques, whereas other institutions place no restrictions on retro-orbital bleeding performed by trained investigators. It is the responsibility of both the investigator and IC ACUC

What anesthetic can I use to bleed rats/mice from the orbital plexus? I am interested in doing retro orbital bleeding of mice/rats for doing FACS analysis of the whole blood. I am looking at a ...

Retro-Orbital Blood Collection Too much blood collected at any one time may cause hypovolemic shock, physiological stress and evendeath. If smaller volumes are collected too frequently, anemia may result. If the retro-orbital route is used for injecting cell suspensions, only single-cell suspensions can be used. Personnel can make a one-cell suspension by using a 70-µm cell strainer (BD Falcon, BD Biosciences, Bedford, MA) to filter the suspension before using it. 1.0 The method, frequency, and volume of blood to be collected must be described in the IACUC protocol. 2.0 If retro-orbital technique is being proposed: 2.1 Include justification for not using a less invasive method of blood collection. 2.2 Include the anesthetic to be used for the procedure. 3.0 If cardiac puncture is being proposed: Training Requirements: Listed below are the training requirements personnel need to complete in order to work with animals at the University of Washington. For a full listing of the Training Program's course offerings, please visit the Online Courses and In Person Courses pages.

Retro-orbital bleeding can be conducted in awake mice. A topical ophthalmic anesthetic should be applied prior to the procedure. Alternatively, systemic anesthesia should be considered if compatible with experimental design. Due to pain and distress issues retro-orbital sampling in the rat is best conducted under general anesthesia. Episodes of retro-orbital bleeding should be every other week only. Repeated bleedings should use alternate eyes. The maximum number of bleeds per animal is two per eye. Animals must be fully anesthetized. A drop of proparicaine topical ophthalmic anesthetic in each eye is recommended to minimize discomfort. Standard heparinized or non- Training Requirements: Listed below are the training requirements personnel need to complete in order to work with animals at the University of Washington. For a full listing of the Training Program's course offerings, please visit the Online Courses and In Person Courses pages.

Due to the nature and anatomy of rat tissues, retro-orbital bleeding may NEVER be performed in an unanesthetized rat. 4. It is best to alternate eyes in successive bleeds. Do not bleed from a damaged (ie. abscessed or ruptured) eye. In the event that one eye is damaged, the second eye can be bled. Retro-Orbital Bleeds (ROB) •The retro-orbital sinus (mouse) and plexus (rat) is a system of dilated venous channels at the back of the orbit. •Blood can be collected from this area in anesthetized animals using a sterile hematocrit tube. •Anesthesia is always required; Isoflurane is recommended.

Dec 13, 2012 · Get YouTube without the ads. Working... Skip trial 1 month free. Find out why Close. Retro orbital mice Srinath Subramanyam. Loading... Unsubscribe from Srinath Subramanyam? The primary method used in the United State is retro-orbital bleeding. This is a rapid and efficient bleeding method but extremely inhumane for the mouse. In fact, the United Kingdom has banned this procedure.

Episodes of retro-orbital bleeding should be every other week only. Repeated bleedings should use alternate eyes. The maximum number of bleeds per animal is two per eye. Animals must be fully anesthetized. A drop of proparicaine topical ophthalmic anesthetic in each eye is recommended to minimize discomfort. Standard heparinized or non- D. Retro-orbital Bleeding Procedure Standard heparinized or non-heparinized micro-hematocrit capillary tubes can be used for blood collection. The animal is held by the back of the neck and the loose skin of the head is tightened with the thumb and middle finger.

In the first installment, we discussed the general blood withdrawal consideration and reviewed the retro-orbital eye bleed, tail snips and nicks, as well as intra-cardiac blood collection methods. Here, we will outline the procedures for blood collection from facial, submandibular, saphenous, and femoral veins. Retro-orbital (RO) bleeding in the rat with recovery should be only used under general anesthesia and only in circumstances where other blood sampling techniques are not feasible (e.g., peripheral veins used for dosing).

Retro-orbital (RO) bleeding in the rat with recovery should be only used under general anesthesia and only in circumstances where other blood sampling techniques are not feasible (e.g., peripheral veins used for dosing).

Oct 20, 2018 · Apply gentle pressure with gauze to stop the bleeding. The bleeding will stop in seconds. Release the mouse that will self-groom. Rotate the side of cheek bleeding for the next time point. For example, we performed the 5 min time point bleeding on the right cheek, the 10 min on the left cheek and the 15 min on the right cheek again. The retro-orbital injection is an acceptable alternative to the tail vein injection route, if technicians receive proper training. The route designated by the protocol should therefore be based on the researcher’s level of training and proficiency with the retro-orbital or tail vein injection.

institutions recommend the use of retro-orbital bleeding only for specific applications and encourage investigators to use other techniques, whereas other institutions place no restrictions on retro-orbital bleeding performed by trained investigators. It is the responsibility of both the investigator and IC ACUC Retro-Orbital Bleeds (ROB) •The retro-orbital sinus (mouse) and plexus (rat) is a system of dilated venous channels at the back of the orbit. •Blood can be collected from this area in anesthetized animals using a sterile hematocrit tube. •Anesthesia is always required; Isoflurane is recommended. retro-orbital healing and minimization of the inflammatory response. o If additional blood is needed, alternate collection sites (saphenous, tail tip, etc.) should be incorporated into the sampling protocol, in consultation with the CAR vet staff. Retro-orbital (RO) bleeding in the rat with recovery should be only used under general anesthesia and only in circumstances where other blood sampling techniques are not feasible (e.g., peripheral veins used for dosing). • Retro-orbital sampling can be used in both mice and rats by penetrating the retro-orbital sinus in mice or plexus in rats with a capillary tube or Pasteur pipette. • Rapid – large number of animals can be bled within a short period of time. • Obtainable volume: medium to large. • Good sample quality. What anesthetic can I use to bleed rats/mice from the orbital plexus? I am interested in doing retro orbital bleeding of mice/rats for doing FACS analysis of the whole blood. I am looking at a ...