Table 1.

Estimated timing for each test for n=10 animals and n=2 operators.

Week #	Test	Comments
1	NOVEL OBJECT RECOGNITION Day 1. Habituation: 30 min/mouse Day 2. Training phase: 5-15 min/mouse Test phase: For STM, after 1 hour of training phase. 10 min/mouse. Day 3. Test phase: For LTM, after 24h of training phase. 10min/mouse.	For Day 1, estimated time is 5-6 hours. For Day 2, estimated time is between 3:30h and 5:15h with STM same day or 2:00h - 3h without STM. For Day 3, estimated time is 2h.
2	MUSCLE STRENGTH AND NEUROMUSCULAR FUNCTION Day 1. Grid Hanging. 3 trials per animal allowing 30 min in between trials. * Day 2. Grip Strength. 3 trials per animal allowing 30 min in between trials. Time estimate is 3 min per animal. Day 2. Frailty index. 15min/animal Day 3. REST Day Day 4. Hanging Bar. 3 trials per animal allowing 30 min in between trials. *** Day 5. Rotarod. 3 trials per animal allowing 30 min in between trials. Time estimate is 3 min per animal.	*depending the age/sex of the animals, estimated total time might vary between 1h and 4h. ** This will take 2-3h *** depending the age/sex of the animals, estimated total time might vary between 1h and 3h. SIDE NOTE: in some tests, total time estimated will depend of the number of different setups available. Some apparatus allow the operator to run multiple animals at a time like multi-channel rotarod or grid hanging test. However the operator must be continually scanning and monitoring all animals during this time.
3	BODY COMPOSITION and ENERGY BALANCE Day 1. Body Composition Test. Experiment takes 2 min/mouse. Estimated time is 1h. Day 2. Energy balance test. Prepare the animals, calibrating and setting up the experiment and run the experiment. Estimated time 2h. Day 3. Energy balance test. The software is up and running recording in vivo metabolic data. * Day 4. Energy balance test. The software is up and running recording in vivo metabolic data. * Day 5. Energy balance test. Stop the experiment, exporting the data, cleaning the equipment and putting animals back in home cages and room. Estimated time 2h.	*schedule based on a 72h experiment set-up with two light cycles and two dark cycles of 12h/each. Side note: During these recording days, animals have to be checked by the operator to make sure they are in good condition and have access to food and water. Also software needs to be checked in order to make sure data is being collected accurately. Two people can expedite the set up of these tests. One person does the body composition measurement, the other sets up the in vivo metabolic machine for calibration and measurement.
4	GLUCOSE TOLERANCE TEST Day 0. Fast animals 6 or 16 hours before the start of the test Day 1. Test day. The last of the time points is at 2 hours after the beginning of the test. To allow for setup and clean up, an additional 1.5-2h is needed. Therefore, total time required is 3.5-4h.	Due to the invasive and stressful character of the test, we let the animals rest the whole week after the test. A skilled person can inject between 20-30 mice within 15min. After the conclusion of the test, animals are returned to their home cage. Mice should be checked after 4-6h to ensure they have returned to normal.
5	INSULIN TOLERANCE TEST Day 1. Fast animals. Animals need to be fasted 3-6h before the experiment.* Test. The last of the time points is at 2 hours after the beginning of the test. To allow for setup and clean up, an additional 1.5-2h is needed. Therefore total time required is 3.5-4h.	*Fasting periods might vary between different studies. We propose for longitudinal studies a fasting between 3h and 6h. Due to the invasive and stressful nature of this test, we let the animals rest the whole week after the test. A skilled person can inject between 20-30 mice within 15min. After the conclusion of the test, animals are returned to their home cage. Mice should be checked after 4-6h to ensure they have returned to normal.
6	CARDIAC FUNCTION Day 1. Echocardiogram. Mice are anesthetized using isoflurane-oxygen Echocardiograms can take up to 30-45min per animal depending on the age, strain and bodyweight. Obese mice can be harder to image given their excess amounts of fat. Day 2. Data processing. 1h	Caution must be taken for old mice who are more vulnerable to death from isoflurane. A skilled operator can process up to 10 animals per day. Mice should be given at least 5 days (longer for aged mice) to recover from the isoflurane.