906
edits
Changes
added ordering details for humulin
*Beaker for weighing mice
*Syringes
*0.1 U/mL humulin in PBS (make as 10uL of 100 U/mL in 10 mL, sterile filtered). This will correspond to 1 U/kg injections. This may need to be adjusted depending on the insulin sensitivity of the mice. This is diluted from Humulin R-100 and is purchased through the veterinary staff.
*Timer
==Protocol==
*Remove food from mice for about 6h. Add do not feed tag to cages, or move cage to procedure room.
*Typically starve the mice at 9AM and aim to start injections at 3 PM
*Weigh mice, mark tails as if necessary and take fasting glucose measurement via a tail clip.
*Prepare glucose syringes with 10 uL per g mouse weight (ie for a 30g mouse, 300 uL).
*At approximately 1 min intervals, inject appropriate amount of glucose insulin into interperitoneal cavity of the mouse.**Immobilize mouse and restrain tail with one hand**Aim needle between the midline and the hip bone**Insert syringe (do not inject) into cavity*Pull up syringe and ensure that only air is aspirated. If urine or blood appears in the syringe, start over*Eject syringe.*At desired intervals (normally 15, 30, 45, 60, 75, 90 , 105 and 120 min), take blood glucose measurements from tail vein. If needed re-snip the tail vein. When measuring glucose just lift the tail of the mouse, while leaving it in the cage, rather than removing and restraining the mouse.
*Analyse data by both % change from fasting glucose and absolute values
