SOP for Lung Function

Table of Contents

1. Purpose

2. Safety Requirements

3. Associated Documents

4. Notes

5. Quality Control

6. Equipment

7. Supplies

8. Procedures

1.Purpose


1.1 : To examine the lung function of a mouse by using unstrained whole body plethysmography.

2. Safety Requirements


2.1 : General laboratory procedures should be followed, which include: no eating, no chewing gum, no drinking, and no applying of cosmetics in the work area. The researcher must wear the laboratory coat, gloves and a mask during the experiment.

3. Associated Documents


3.1 :  http://www.buxco.com/products_technologies_wholebodyplethysmography.aspx

4. Notes


4.1 : The mice must be maintained in a controlled environment with stable temperature, humidity, and air pressure. Keep minimal disturbing and stress on the tested mice whenever is possible.
4.2 : The lung function of a mouse is age/sex/strain dependent. It is important to have the age-matched mice with the same sex and strain background in a single experiment.
4.3 :The measurement of mouse lung function is airflow and volume dependent, so we should control atmospheric pressure stable during the experiment.
4.4 :The Buxco chambers will be cleaned after all cases of a single user are completed.

5. Quality Control


5.1 : Calibrate the whole body plethysmograph system before measurement.
5.2
: Before the test, the mouse will habituate in the Buxco chamber for 15min.
5.3 : We will avoid opening the lab door to maintain the atmospheric pressure stable.

6. Equipment


6.1 : Unrestrained whole body plethysmograph (Buxco) chamber for mouse
6.2 : Ponemah (signal recorder;preamplifier)
6.3 : Pressure transducer
6.4 :
Bias flow regulator
6.5 : FinePointe (lung function analysis software for mouse)
6.6 : Aerosol system for whole body plethysmograph

7. Supplies


7.1 : Cap
7.2 : Dry stone
7.3 : Gloves
7.4 : Mask
7.5 : Ethanol 70%
7.6 : paper towel
7.7 : PBS
7.8 : Methacholine
7.9 : HOCl

8. Procedures


8.1 : Turn on the computer.
8.2 : Turn on the bias flow regulator, and link the tube to the Buxco chamber.
8.3 : Turn on the preamplifier, and link the transducer to the Buxco chamber.
8.4 : Calibrate the Buxco system with FinPointe software.
8.5 : Put the mouse in the Buxco chamber to measure lung function.
8.6 : The mouse is allowed to habituate in the Buxco chamber for 15min.
8.7 : Spray aerosol of PBS or Methacholine for 3min (user can adjust the time).

8.8 : Record respiratory data for 3min (user can adjust the recording time).
8.9 : Allow a 5-min resting (or longer, depending on the mouse respiration condition) before next procedure.
8.10 : Repeat steps 7-9. The order of neutralization is PBS、Methacholine 3.125、12.5、25、50 mg/ml for mouse inhalation (up to 4 different Methacholine concentrations).
8.11 : Move the mice to their home cage and then clean the Buxco chamber.

 

P.S.
The user can choose to skip or revise the steps of 7~10.
If the user wants to test the basal lung function, we propose to skip the steps of 7~10. The basal respiratory parameters will be measured for 1 hour and spray a single dose of aerosolized Methacholine (12.5 mg/ml) at the 50th min.
The researcher should avoid inhaling the aerosolized Methacholine, because it would induce asthma.