Difference between revisions of "Insulin Tolerance Test"
From Bridges Lab Protocols
Davebridges (Talk | contribs) (initial page) |
Davebridges (Talk | contribs) m (fixed some formatting errors) |
||
| Line 7: | Line 7: | ||
*0.07U/mL humulin in PBS (make as 70uL of 100 U/mL in 10 mL, sterile filtered). This will correspond to 0.7mU/kg injections. | *0.07U/mL humulin in PBS (make as 70uL of 100 U/mL in 10 mL, sterile filtered). This will correspond to 0.7mU/kg injections. | ||
Timer | Timer | ||
| − | |||
| + | ==Protocol== | ||
*Remove food from mice for about 6h. Add do not feed tag to cages, or move cage to procedure room. | *Remove food from mice for about 6h. Add do not feed tag to cages, or move cage to procedure room. | ||
*Bring mice to procedure room. Put water bottle on cage. | *Bring mice to procedure room. Put water bottle on cage. | ||
Revision as of 19:49, 2 September 2010
Materials (Bring Downstairs
- Glucometer - AccuChek Advantage
- Glucose Test Strips - AccuChek Comfort Curve. Order these through Materiels Service by calling 936-6077 and ordering product 2535. For information see [here]
- Scale
- Beaker for weighing mice
- Syringes
- 0.07U/mL humulin in PBS (make as 70uL of 100 U/mL in 10 mL, sterile filtered). This will correspond to 0.7mU/kg injections.
Timer
Protocol
- Remove food from mice for about 6h. Add do not feed tag to cages, or move cage to procedure room.
- Bring mice to procedure room. Put water bottle on cage.
- Weigh mice, mark tails as necessary and take fasting glucose measurement.
- Prepare glucose syringes with 10 uL per g mouse weight (ie for a 30g mouse, 300 uL).
- At 1 min intervals, inject appropriate amount of glucose 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 and 120 min), take blood glucose measurements from tail vein
- Analyse data by both % change from fasting glucose and absolute values
