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TSE Metabolic Cages

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TSE Metabolic Cages

The TSE Calorimetry System includes the capacity for measuring O2 consumption, CO2 production, food intake, water intake, air temperature, physical activity in x, y, and z planes and running wheel activity on 16 mice or rats simultaneously. This system is maintained by East Carolina Diabetes and Obesity Institute (ECDOI) and is open for use to any ECU faculty member. ECDOI is responsible for oversight of the system. Please contact Jim Aloor for scheduling, calibration, and training on the system. You must receive training prior to using the system. The Standard Operating Procedures (SOP) below must be followed.

Standard Operating Procedure

  1. The system represents a new housing environment for rodents. Animals must be single housed to obtain valid data for all metabolic parameters measured. Enrichment may be included in cages if physical activity is not being monitored. If activity data is required, enrichment objects should not be added to insure infrared activity system accurately detects all animal movement.
  2. All investigators must have “metabolic cage housing” listed in their existing AUP prior to use; otherwise, an amendment must be filed to an existing AUP. Please indicate the dates and the total number of animals (broken down by treatment groups) that will be housed in the metabolic cages. Also indicate that you will follow the SOP and attach a copy of the SOP (this memo) in the amendment to the IACUC.
  3. All investigators must notify the Facility Manager prior to initiating housing in this system. Since this is a new environment for rodents, it is strongly recommended that investigators allow at least 48 h acclimation prior to initiating data collection. Data may be collected for an additional 24-72h.
  4. While using the TSE system, the investigator/investigator’s personnel will be responsible for checking the animals at least twice per day. The investigator will also be responsible for replenishing/replacing all food, water and bedding as needed and insuring that the system is functioning properly. No maintenance will be provided by the animal care staff.
  5. The TSE System is on red-line power and is backed up by an uninterrupted power source (UPS) to insure oxygen supply is maintained in the event of an emergency. If there is a power failure and the metabolic cages are in use, the Director of ECDOI (or designated faculty member within ECDOI) will be notified by Animal Care staff. The PI will be notified immediately by the Director or designated ECDOI faculty member.
  6. The investigator and/or investigator’s personnel are responsible, under ECDOI faculty supervision, for assembly/disassembly and cleaning of the system. Parts of the system are delicate and expensive; please exercise extreme caution! Make sure o-rings on feeding/drinking sensors have a thin layer of silicone grease and are in tracks. Use a gentle twisting motion while pushing straight down or pulling straight up – do not angle or torque sensor or the plastic housing will crack.

Cleaning

  • Water Bottles and Drinking Nipples – glass bottles should be sprayed down with Quatricide PV 15 (spray bottle in room), rinsed thoroughly with warm water, and air dried on racks. BE CAREFUL! The screw caps and the drinking nipple components can be dismantled into their individual components, sprayed with Quatriacide 15, rinsed with warm water and air dried.
  • Feeding Baskets – spray down with Quatricide PV 15 and rinse with warm (not hot) water and air dry.
  • Plastic Cage tops – disinfect underside of cage top by wiping down with a soft, lint-free cloth sprayed with Quatricide PV 15 followed quickly by a second cloth moistened with water only to rinse disinfectant from plastic. The plastic is susceptible to cracking/damage from organic solvents, detergents, or alcohol.
  • Cage Bottoms - will be cleaned using standard cagewashing procedures by Animal Care personnel. Cage bottoms may be autoclaved if desired.
  • Activity Sensor Frames – Any bedding/feces should be cleaned from area. The sensor frames should be wiped off with a soft, lint-free cloth moistened with water.
  • Running Wheels – Wheel must be dismantled from housing. Contact director or designated ECDOI faculty for instructions. The wheel and its metal housing should be sprayed with Quatricide PV 15, rinsed with warm running water, and allowed to air dry.
  • Drinking/Feeding Sensors – may be wiped down with a soft lint-free cloth moistened with warm water only. On drinking/feeding sensors, do not pull on the hook or press the hook into the sensor housing.
  • Air drying unit – empty glass reservoir for air drying unit after every run.

Calibration

Gas Calibration should be performed at least once every month or when switch user.

  1. Warm up the system for at least 2hr. Switch on power and load software.
  2. Open Calo value window, and keep it open to observe the sample flow rate 0.2 L/m during the calibration.
  3. Open the calibration program.
  4. Turn on the Low O2 calibration gas tank and adjust pressure.
  5. Disconnect the Sample Out Tube from the Air Drying Unit in the front.
  6. Connect the Sample Out Tube to the Low O2 calibration gas line.
  7. Enter the O2 value from the Gas bottle (see certification, should be 20.00) in the first left field of the calibration window
  8. Check the field auto, set the smoothing to 600 sec. Wait until the buffer is filled to 100% (around 10 minutes). Also check from time to time the sample flow rate in the left window (0.25 L/m).
  9. After the buffer is filled to 100%, uncheck Auto, click Measure 5 times, and then Apply ADC for the first value
  10. Enter in the second left field the higher O2 level (should be 20.90) from the second calibration gas bottle.
  11. Close the valve of the Low O2 gas tank. Open the valve of the High O2 gas tank and adjust pressure.
  12. Check the sample flow rate from the monitor.
  13. Click Auto and wait again until the buffer is 100% filled (check the sample flow from time to time).
  14. After the buffer is 100% filled, uncheck Auto, click measure 5 times, and click Apply ADC.
  15. Click Calculate and Click Save.
  16. Go to CO2 calibration by clicking + and enter the CO2 value (0.050) from this gas tank in the left first field of the window. The High O2 gas should still be connected and flowing.
  17. Check Auto, and set the smoothing to 120 sec.
  18. After the buffer is filled to 100%, uncheck Auto, click Measure 5 times, and click Apply ADC.
  19. Enter the higher CO2 value (0.9500) in the second left field of the window. Close the valve of the High O2 gas and open the valve of the Low O2 gas tank and connect the Sample Out Tube to Low O2 gas line.
  20. Set the smoothing to 600 sec
  21. After the buffer is filled to 100%, uncheck Auto, click Measure 5 times, and click Apply ADC.
  22. Click Calculate and Click Save.
  23. Close the valves on all gas tank and reconnect the sample out tube to air drying unit
  24. After 5 minutes, check the O2 and CO2 values in the Status calo Window. O2 should be around 20.8% and CO2 should be around 0.05% in the reference air.
  25. Ready to put the animals in.

System Feeding & Drinking Calibration should be performed at least once every month or when switch user.

  1. Hang the empty food and drink bottle under the sensors that need to be calibrated.
  2. Click Batch calibration
  3. Click Weight, input 0 and 38.5443 (when use #3 weight), click OK
  4. Click calibration, click boxes that need to be calibrated.
  5. Smooth (30), max drift (5), max variant (30)
  6. Check auto catch
  7. Click clear buffer
  8. After reach 100% buffer, all box will show a figure of 1
  9. Hang #3 weight (in a plastic bag in the cabinet) in one of drinking or feeding bottles.
  10. Click clean buffer.
  11. After reach 100% buffer, the sensor for that box will show a figure of V automatically.
  12. Move #3 weight to the next feeding or drinking bottles, repeat step 9-11 until all required sensors show V.
  13. Click Calculate
  14. Click Save
  15. Close this window. Ready to go.