Pyruvate Tolerance Test: Difference between revisions
Created page with "'''Materials''' Glucometer - AccuChek Advantage Glucose Test Strips - AccuChek Comfort Curve. Order these through Materiels Service by calling 936-6077 and ordering product 2..." |
No edit summary |
||
| Line 1: | Line 1: | ||
'''Materials''' | '''Materials''' | ||
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] | ||
Scale | Scale | ||
Beaker for weighing mice | Beaker for weighing mice | ||
Syringes | Syringes | ||
2.5g pyruvate in 10 mL PBS- This will correspond to 2.5 g/kg injections. | 2.5g pyruvate in 10 mL PBS- This will correspond to 2.5 g/kg injections. | ||
Timer | Timer | ||
'''Protocol''' | '''Protocol''' | ||
Remove food from mice for about 6h. | Remove food from mice for about 6h. | ||
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 if necessary and [[take fasting glucose measurement]] via a tail clip. | 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). | 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. | At approximately 1 min intervals, inject appropriate amount of pyruvate 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, 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. | Analyse data by both % change from fasting glucose and absolute values. | ||