ECG Screening Protocol
Non-invasively Recording Electrocardiograms in Conscious Mice
Revision v2.0 LJ, RVS 20050125
System used: AnonyMOUSE ECG Screening System
Mouse Specifics, Inc. 28 State St., Suite 1112
Boston, MA 02109
1. Turn on the Bio Amplifier ***The Bio Amplifier must be turned on first***
2. Power up computer.
3. Set up Tower: (Refer to diagrams 1 – 3)
- Slide platform on top cap of tower. The open end of the platform should be centered to the arrow on the cap.
- Place 3 electrodes on the plastic lead plate*, which has 3 holes. Connect the lead plate to the platform with the off set hole towards the back.
- Insert the side shields into the slots on the platform. Place the back shield into the channels along the top of the side shields. Use the closest channel to the platform for small to medium sized mice, and one channel back for larger mice. Note: A piece of paper towel is placed beneath the back shield and on each side of the platform to protect it from soiling.
4. Acclimate the Mice:
- Place two mice on the sides of the platform to acclimate. A 10 minute Acclimatization period is recommended before recording data.
- Note: The acclimation period is very important. Body tremors, shivering, and general activity, all perhaps heightened in the new environment, can all introduce electrical signals that confound acquisition of ECG. As the mouse relaxes, it is more likely to contact the electrode pads which allows for a recordable ECG signal.
5. Set up Data Folders:
- Click on Todays EKG Data
- Go to file, then new folder
- Name new folder (example: G3’s mm-dd-yy, STRAINS mm-dd-yy, HLB Mice mm-dd-yy, CONTROLS mm-dd-yy).
- Note: If running more than one type of mouse, i.e. G3, Strain, HLB, you may want to make separate folders for organization)
6. Initialize Chart 4 software and General Set Up:
- Open Chart 4 through the Shortcut Icon on the Desktop
- Several Screens will show up, using your mouse, drag the top screens lower boundary so that channel 1 fills the entire screen.
- Click on top drop down button (upper right corner)
- Scroll down to 2K and click to set.
- Click on next drop down button (under top button)
- Scroll down to 1mv and click to set.
- Click on Channel 1 drop down button.
- Scroll to Bio Amplifier and select
- Bio Amplifier screen will appear. Scroll and set each of the following.
Click 60 Hz Notch
High Pass 0.1 Hz
Low Pass 100 Hz Click OK
6. Initialize Chart 4 Software and General Setup Continued:
-Click on the icon under the word Macro it looks similar to (x =) it is in the Upper left corner
-A Channel 1 Mean screen will appear.
- click on gray area.
- Data Pad Column A Setup screen will appear
- scroll down to Maximum-Minimum and click OK.
- On the icon that looks like a little mountain (Bottom right corner by start button) click once to 2:1 setting
- Enter mice ID# in the log book.
Note: To avoid confusion I enter the mice ID# in order as they are on the pen cards, this helps prevent having two mice of the same sex and ear notch on the tower at the same time. (This will vary based on individual lab procedures for animal marking.)
7. Data Acquisition: Ready to Start
- In order as recorded, Gently place one of the mice that have been acclimating on the platform to the lead plate.
- Click Start and wait and watch for ECG signal to manifest.
NOTE:: while waiting for signal to appear Gently place next
mouse on vacant side of platform to acclimate.
- Once a good 2 to 3 second signal is obtained Press Stop and remove the mouse.
- Block out a 2 second section of ECG signal.
- Go to space bar at bottom of screen and scroll back and forth to find the 2 second section of signal you want to save.
- Select a starting point and highlight by dragging across the screen until you reach the end of the 2 second section.
- Go to File, Save Selection
NOTE: (For the first mouse you run in a group you have to go to Save In: scroll down to Local Disk C, then select EKG Data, then choose the folder in which corresponds to the type of mouse you are working with.
- Once folder is chosen:
- Go to Save as Type and select Chart Text File (*.txt)
- Click on: File name and Type in mouse ID# (62454-MG3)
- Click Save
- On the Pop-Up screen ‘Save Selection As Text’, Click OK
-You are now ready to run your next mouse.
-When you are done running mice close out of the program.
NOTE: Document 1 screen will appear warning you that the document has been changed. Do you wish to save changes? Press→ No
8. Reviewing Data:
- Start up Auto-eMouse (top right corner).
-You will see:
Initializing- - -Please Wait (takes a few seconds)
Auto-eMouse 2.11 Mouse Specifics, Inc. screen.
- Go to User and Type in initials LMJ)(Frequency should be set at ◙ 2000 Hz)
- Select My Computer
- Select Local Disk C
- Select EKG Folder
- Choose folder to Review, Click OK
- Highlight all Mice in the folder, Click GO
- The first ECG Signal (for example 62454-MG3.txt) appears.
- Go to the Minus button, click it, a Red Dot will appear on one of the peaks.
- Keep selecting the Minus Button until all the peaks have a Red Dot.
(NOTE: The last peak will NEVER have a Red Dot)
- Once finished placing the Red Dots:
- Click Go
- A new screen “Final & Processed, ECG Signal for Data Files
appears, it is the Trace of ECG signal showing P, Q, R, S, T
points.
- Press Save
- A window “File to Write” will appear.
- For the First mouse you do in a group you have to:
-Click in File name.
- type in EKGmmddyy (this creates a result folder)-Click Save
-Close (Next signal will appear)
-Repeat Select for Red Dots
-Select Save
- After the First mouse and the window “Select File to write” appears select the results folder you just created. Repeat for every mouse data set.
- When finished, Exit, a Warning Screen appears Select Yes and then Close
- Once all your data is processed in Auto emouse: Left click→ Exit
- Close out of screen.
- Shut down computer and Bio Amplifier Box.
9. Clean Up:
- Disassemble tower
- remove side shields
- unsnap lead plate
- peel off electrodes (Do not throw plastic lead plate away)
- Remove platform
- Wash shields, platform, lead plate, and forceps with mild soap and a Teri Wipe cloth.
9. Clean Up Continued:
- Rinse out Teri Wipe and wipe down tower and desk. (Do not wipe the snaps on top of the cap of the tower)
- Wipe desk and teacart down with Alcohol.
- Now you are ready for another day
* Lead plates formal Name and Ordering information follows:
Snap-Style Pre-Gelled Plates
Source One Health Care Tech Inc.
8020 Tyler Blvd.
Mentor OH 44060
ECG SETUP
Diagram 1
Diagram 2
Diagram 3
Tower with just the platform Tower assembled
