In vivo phenotyping
Objective: Assessment of physiological and behavioral characteristics in living mice of different ages
Gather these items before starting the experiment. Check off items as you prepare.
Equipment1
not specified • not mentioned • not mentioned • not mentioned
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Protocol Steps
Acquire mice
Obtain C57BL/6J mice of two age groups from vendor
Note: Two cohorts: 13 weeks old (young) and 93 weeks old (old)
View evidence from paper
“C57BL/6J mice of either 13 weeks old ("young") or 93 weeks old ("old") were purchased from Janvier (St. Berthevin, France)”
House mice
Place mice in group housing conditions
Note: Mice remain group housed throughout the study period
View evidence from paper
“Mice were group housed and received standard chow”
Provide standard diet
Feed mice standard chow with specified macronutrient composition
Note: Diet catalog #2018 contains 18% protein, 50% carbohydrate, 6.0% fat
View evidence from paper
“received standard chow (#2018, containing 18% protein, 50% carbohydrate and 6.0% fat; Harlan Laboratories, Madison, WI, USA)”
Conduct in vivo phenotyping
Perform assessment of physiological and behavioral characteristics in living mice
Note: Specific phenotyping methods not detailed in this methods section
View evidence from paper
“After in vivo phenotyping, mice were sacrificed after overnight fasting”
Fast mice overnight
Withhold food from mice for overnight period prior to sacrifice
Note: Fasting occurs before sacrifice procedure
View evidence from paper
“mice were sacrificed after overnight fasting, reaching the age of 24 or 103 weeks”
Sacrifice mice
Euthanize mice at specified ages
Note: Young mice sacrificed at 24 weeks; old mice sacrificed at 103 weeks
View evidence from paper
“mice were sacrificed after overnight fasting, reaching the age of 24 or 103 weeks”
Collect heparinized plasma
Obtain blood samples in heparinized tubes
Note: Collected immediately after sacrifice
View evidence from paper
“Heparinized plasma was taken and tissues were frozen in liquid nitrogen”
Freeze tissue samples
Preserve tissue samples in liquid nitrogen for later biochemical and molecular analyses
Note: Tissues frozen immediately after collection
View evidence from paper
“tissues were frozen in liquid nitrogen for biochemical and molecular analyses”