Difference between revisions of "Pyruvate Tolerance Test"
From Bridges Lab Protocols
Davebridges (Talk | contribs) (Updated layout of the protocol) |
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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.
