Pyruvate Tolerance Test: Difference between revisions
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==Materials== | |||
* 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 | |||
* 2.5g pyruvate in 10 mL PBS- This will correspond to 2.5 g/kg injections. | |||
* Timer | |||
==Protocol== | |||
* Remove food from mice for about 6h. | |||
* 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 pyruvate syringes with 10 uL per g mouse weight (ie for a 30g mouse, 300 uL). | |||
* At approximately 1 min intervals, inject appropriate amount of pyruvate 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. | |||
Remove food from mice for about 6h. | |||
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 pyruvate syringes with 10 uL per g mouse weight (ie for a 30g mouse, 300 uL). | |||
At approximately 1 min intervals, inject appropriate amount of pyruvate 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. | |||
Revision as of 16:35, 27 April 2017
Materials
- 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
- 2.5g pyruvate in 10 mL PBS- This will correspond to 2.5 g/kg injections.
- Timer
Protocol
- Remove food from mice for about 6h.
- 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 pyruvate syringes with 10 uL per g mouse weight (ie for a 30g mouse, 300 uL).
- At approximately 1 min intervals, inject appropriate amount of pyruvate 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.