Optical mapping

1. Clean the working area, it's almost certainly a mess.
2. Ensure bath chamber is surrounded by 6 LEDs and camera/arm is attached to the table and above the chamber.
3. Turn on the LED power supply (blue box), the switch is on the back. On the front side, turn the lower dial to page 4 (test) and turn on the red LEDs.
4. Adjust the position of LEDs to optimize the illumination of the chamber while being careful not to obstruct the camera's view or mobility.
5. Repeat for green LEDs.
6. Use the two upper knobs to increase the current to LEDs.
7. Use the lower knob of LED power supply to turn to page 3, external pacing. Ensure that the cable from camera frame readout is connected to the input port.
8. Ensure pacing apparatus is set up according to chosen methods ( see section on pacing ).
9. Choose a camera lens and place the long pass filters on the lens before attaching to camera. The direction of filters is important. they have little arrows on the side which indicate the direction light should pass through the lens.
10. Place prepared heart in the chamber ( see section on heart preparation ).
11. Turn on the camera.
12. At the computer, open RSI image software (blue icon at the right side of desktop.
13. At the lower right corner of the window, click focus. Exit the program, no need to save the focus.
14. Open the folder at the bottom right of screen named MultiRecorder. Scroll to the bottom and run the program qtMultiRec.exe.
15. Adjust the size of the window, but don't click the maximize button. This program doesn't like that.
16. Click the camera button in the upper left corner of window. Choose a zoom of 200%.
17. Adjust exposure time (1ms is usually good) and gain.
18. Use the lens and arm to focus on the heart.
19. Click traces button at the upper left. At the bottom a new window will appear. At the bottom of this window, click options. If doing both calcium and voltage, uncheck the option "show odd and even frames together" and check "show odd and even frames separate." Adjust the time in the input box to something on the order of 5 seconds.
20. Go to file>settings>recording and choose a folder to save data to. Create a folder in your chosen location and select it.
21. Begin your pacing protocol and enter a comment about the pacing interval for each new recording ( i.e., 500ms).
22. Choose your recording period. For open ended, you stop the recording when you wish to.
23. Click record. The data automatically saves to your selected folder in a convenient format. There are inputs at the top left of the window labelled counter 1 and counter 2 that allow you to alter the experiment number and recording number if you wish ( the recording number automatically increases by 1 each time you take another recording and these counters generally don't have to be touched ).