Difference between revisions of "Insulin Tolerance Test"
From Bridges Lab Protocols
Davebridges (Talk | contribs) (updated with new starvation and added additional information) |
Davebridges (Talk | contribs) (added ordering details for humulin) |
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*Beaker for weighing mice | *Beaker for weighing mice | ||
*Syringes | *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 | + | *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 | *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. | ||
| − | |||
*Typically starve the mice at 9AM and aim to start injections at 3 PM | *Typically starve the mice at 9AM and aim to start injections at 3 PM | ||
| − | *Weigh mice, mark tails | + | *Weigh mice, mark tails 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). | *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 | + | *At approximately 1 min intervals, inject appropriate amount of insulin 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 |
| − | *Insert syringe (do not inject) into cavity | + | **Insert syringe (do not inject) into cavity |
| − | * | + | **Eject syringe. |
| − | *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. |
| − | *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 | *Analyse data by both % change from fasting glucose and absolute values | ||
Revision as of 15:29, 17 January 2014
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. 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 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 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
- 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
