Difference between revisions of "Pyruvate Tolerance Test"
(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...") |
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'''Materials''' | '''Materials''' | ||
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Glucometer - AccuChek Advantage | Glucometer - AccuChek Advantage | ||
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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] | ||
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Scale | Scale | ||
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Beaker for weighing mice | Beaker for weighing mice | ||
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Syringes | Syringes | ||
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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. | ||
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Timer | Timer | ||
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'''Protocol''' | '''Protocol''' | ||
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Remove food from mice for about 6h. | Remove food from mice for about 6h. | ||
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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 | ||
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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. | ||
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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). | ||
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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. | ||
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Immobilize mouse and restrain tail with one hand | Immobilize mouse and restrain tail with one hand | ||
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Aim needle between the midline and the hip bone | Aim needle between the midline and the hip bone | ||
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Insert syringe (do not inject) into cavity | Insert syringe (do not inject) into cavity | ||
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Eject syringe. | Eject syringe. | ||
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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. | ||
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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 19:40, 23 June 2015
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.
