Difference between revisions of "Insulin Tolerance Test"
From Bridges Lab Protocols
Davebridges (Talk | contribs) (updated insulin concentration) |
Davebridges (Talk | contribs) (updated with new starvation and added additional information) |
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| − | ==Materials (Bring Downstairs== | + | ==Materials (Bring Downstairs)== |
*Glucometer - AccuChek Advantage | *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] | *Glucose Test Strips - AccuChek Comfort Curve. Order these through Materiels Service by calling 936-6077 and ordering product 2535. For information see [here] | ||
| Line 5: | Line 5: | ||
*Beaker for weighing mice | *Beaker for weighing mice | ||
*Syringes | *Syringes | ||
| − | *0. | + | *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 |
| − | Timer | + | *Timer |
==Protocol== | ==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. |
| − | * | + | *Separate mice into individual cages and mark with mouse ID. Put a water bottle on the cage. |
| − | *Weigh mice, mark tails as necessary and take fasting glucose measurement. | + | *Typically starve the mice at 9AM and aim to start injections at 3 PM |
| + | *Weigh mice, mark tails as 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). | *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. | + | *At approximately 1 min intervals, inject appropriate amount of glucose into interperitoneal cavity of the mouse. |
*Immobilize mouse and restrain tail with one hand | *Immobilize mouse and restrain tail with one hand | ||
*Aim needle between the midline and the hip bone | *Aim needle between the midline and the hip bone | ||
Revision as of 15:00, 13 May 2011
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
- 0.1 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
- Timer
Protocol
- Remove food from mice for about 6h. Add do not feed tag to cages, or move cage to procedure room.
- Separate mice into individual cages and mark with mouse ID. Put a water bottle on the cage.
- Typically starve the mice at 9AM and aim to start injections at 3 PM
- Weigh mice, mark tails as 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 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
