Dec 13, 2022
Advanced Dynamic Weight Bearing system for mice
- 1MD,MS
Protocol Citation: Fanglin Lu 2022. Advanced Dynamic Weight Bearing system for mice. protocols.io https://dx.doi.org/10.17504/protocols.io.eq2ly76pmlx9/v1
Manuscript citation:
Lu F, Kato J, Toramaru T, Sugai M, Zhang M, Morisaki H. Objective and Quantitative Evaluation of Spontaneous Pain-Like Behaviors Using Dynamic Weight-Bearing System in Mouse Models of Postsurgical Pain. J Pain Res. 2022 Jun 2;15:1601-1612. doi: 10.2147/JPR.S359220. eCollection 2022.
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: December 10, 2022
Last Modified: December 13, 2022
Protocol Integer ID: 73823
Keywords: spontaneous pain-like behavior, pain evaluation
Abstract
We can assess spontaneous limb pain-like behaviors using advanced DWB apparatus, wherein changes in the postural equilibrium of each freely moving animal were tracked and analyzed.
This protocol outlines the procedures from calibration to data analysis.
Materials
PC
Advanced DWB apparatus (Bioseb, Vitrolles, Provence-Alpes-Côte D’Azur, France; catalog number: BIO-DWB-M)
We made the apparatus a hat to reduce the reflection of room light.
3000mL water
Before start
Install the DWB software set (drivers for digital USB camera, driver for PC/sensor interface, video codec, advanced DWB software).
Calibration
Calibration
Note
A calibration must be performed before experiment.
Connect the sensor and the camera to the PC.
5m
Lauch the software and select the sensor type "Mice".
Click "Full calibration".
Enter the calibration weight, then click "Next".
For mice, 2000-3000g is preferred, and 7000-10000g for rats
Adjust the display angle of the sensor to match the real sensor direction by clicking "<" or ">" on the top.
Make sure no weight is on the sensor and click "Tare".
Click "Gain preview", and apply the calibration weight (3000g) indirectly on the sensor.
Follow the guidance on the right.
Acquisition setting
Acquisition setting
Embed the sensor in the floor of the chamber.
Click "New" to create a new experiment project.
Click "Browse..." and select the calibration file that is previously saved.
Click "Tare and Mask".
Follow the guidance on the right.
Click "Next".
Click "Video setting" to make sure that the DWB software is connected to the digital camera, not WebCam of the PC.
Click "OK".
Data acquisition
Data acquisition
Place the mouse in the chamber.
Adjust the camera aperture and focus to get a bright and clear image.
Click "Capture" to lauch the acquisition.
Input the weight and click "OK".
Note
We suggest to tick " Experiment automation" to schedule the acquisition automatically.
Latency = 60ses: 1min for acclimatization before the recording.
Record time= 5min: We record 5min for each mouse.
The mouse is allowed to move freely in the chamber and changes in postural equilibrium were synchronously and automatically tracked and analyzed by the software.
2m
When the capturing is over, click "analysis" and compress the data file.
Place the mouse back to its cage and clean the chamber before the placement of next mouse.
1m
Data analysis
Data analysis
8m
8m
Click "Menu"→"Open experiment file..."
Manually validate each automatically presumed paw position to avoid error identification afterward.
Tick "Independent paws setting" and "Go through validation".
We suggest to choose "never" make a backup during validation as it takes time to save the file.
Click "Legend" to help validating each paw position.
Note
A zone was considered valid when the following parameters were detected: ≥0.8 g on one captor with a minimum of two adjacent captors recording ≥1.0 g. A time segment was considered valid if ≥3 stable pictures were detected.
6m
After validation, click "Result".
Click "Graphics" to visualize the results.