小鼠常用实验技术课件.ppt

1、Laboratory Animal Handling Technique -Mouse-Rat-RabbitObjectivenTo comply with the Animal Welfare Ordinance and avoid mishandling of animal in research nTo provide basic concepts of animal handling technique to new animal usernWhile offering our concept and techniques to our animal user,we also enco

2、urage comments from experienced animal users.By doing so,we would enrich our knowledge in the field of laboratory animal research on both sides and further benefit animal welfare as well as the credibility of research result in our universityLaboratory Animal Handling Technique-MousenA.Blood collect

3、ion from tail veinnB.Blood collection from orbital sinusnC.Blood collection from cardiac puncturenD.Blood collection from saphenous veinnE.Intraperitoneal injectionnF.Subcutaneous injectionnG.Oral FeedingnH.SexingBlood Collection From Tail in MousenFor collection of small amount of blood(Approximate

4、 0.1 ml)Tools for Blood Collection from Tail n75%alcohol cotton ball for surface disinfectionnSmall plastic bottle with 1/2 cm diameter holes in both ends as mouse restrainernScissorsnPipetteman and tipsnA vial for blood collectionPlacing a mouse on a cage lid and grasping the loose skin behind the

5、ears by the thumb and forefingerPush the mouse into the restrainerLeave the tail of the mouse outside the cover of the restrainerAmputate the tip of the mouse tail by scissorsMassage the tail and collect blood by pipettemanBlood Collection From Orbital Sinus in MousenShould apply anesthetic before b

6、lood withdrawnA convenience and easy apply method for blood collection in mousenCollect amount up to 0.5 mlTools for Blood Collection from Orbital Sinus in Mousen75%alcohol cotton ball for surface disinfectionnHypnorm for general anestheticn27 G needle with 1 ml syringe for injectionnGlass capillary

7、 tube and vial for blood collectionAnesthetize a mouse by intraperitoneal injection of HypnormUse a sharp end glass capillary tube to penetrate the orbital conjunctiva and rupture the orbital sinusCollect blood with a vialBlood Collection From Cardiac Puncture in MousenFor collect up to 1 ml of bloo

8、d within a short period of timenMust be performed under general anestheticTools for Cardiac puncture in Mousen75%alcohol cotton ball for surface disinfectionnHypnorm used as anestheticn27G needle with 1 ml syringe for injectionn24G needle with 3 ml syringe for blood withdrawAnesthetize a mouse by in

9、traperitoneal injection of HypnormDisinfect the thorax area with 75%alcohol cotton ballSearch for the maximum heart palpitation with your fingerInsert a 24G 1”needle through the thoracic wall at the point of maximum heart palpitationWithdraw blood slowly by your right handBlood Collection From Saphe

10、nous Vein in MousenThis method is used of multiple samples are taken in the course of a daynIt can also be applied on rats,hamsters,gerbils and guinea-pigsTools for blood collection from Saphenous vein in micen75%alcohol cotton ball for surface disinfectionn50 ml syringe tube with small holes at the

11、 end as restrainerna scalpel and shaver for remove of hair n24 G 1“needle for release of bloodntips and pipetteman for blood collectionPlacing a mouse on a cage lid and grasping the loose skin behind the ears with your thumb and forefingerPlace the mouse in the restainerPull out the leg and removed

12、the hair by a assistantHair can also be shaved by using a small scalpelThe saphenous vein is seen on the surface of the thighApply vaseline after disinfect the surface area to reduce clotting and coagulation during blood collection.Use a 24 G 1”needle to puncture the vein and release blood from the

13、saphenous veinUse a Microvette or a pipetteman with tip to collect blood from the saphenous veinApproximate 100 microliters can be collectedFlex the foot of the mouse to reduce the flow of blood back to the puncture siteA cotton ball is applied to the puncture site to stop further bleedingIntraperit

14、oneal Injection in MousenA common method of administering drugs to rodentsTools for Intraperitoneal Injection in Mousen75%alcohol cotton ball for surface disinfectionn25G 1/2”needle with 1 ml syringe for injectionPlace a mouse on a cage lid and grasping the loose skin behind the ears with your thumb

15、 and forefingerAs soon as the mouses head is restrained,the mouse can be picked up and the tail secured within your ring finger and little fingerThe injection site should be in the lower left quadrant of the abdomen because vital organs are absent from this area.Only the tip of the needle should pen

16、etrate the abdominal wall to prevent injection into the intestine.Subcutaneous Injection in MousenThe most common method for immunology studiesTools for Subcutaneous Injection in Mousen75%alcohol cotton ball for surface disinfectionn25G 1“needle with 1 ml syringe for injectionPick up a nude mouse an

17、d spin its tail to put it in a faint conditionGrasp the loose skin on the back of the mouse from ears along the legs and restrain the legs with your ring finger and little fingerAfter disinfect the surface area,insert the needle in the lateral side of the abdominal wall and push upwards to the armpi

18、t of the mouseInject the substance slowlyA lump of injection substance can be seen through the skin after injectionOral Feeding in MousenGastric intubation ensures that all the material was administerednFeeding amount limited to 1%of body weightTools for Oral Feeding in MousenA 18 G stainless steel,

19、ball tipped needlena gloveGrasp the loose skin on the back of the mouse and restrain its tail with your ring finger and little finger.Then,introduce the feeding tube from the pharynx in to the esophagus when the mouse is in the act of swallowing.Common complications associated with gastric intubatio

20、n are damage to the esophagus and administration of substance into the trachea.Careful and gentle passage of the feeding needle will greatly reduce these possibilities.The anatomy picture showed the position of the feeding needle tip inside the esophagus with the heart and sternum removed.Sexing mice-The distance between the anal and genital orifices is greater in the male(left)compared to the female(right).