This article describes a high temporal and spatial resolution technique to optically image action potential movement on the surface of Langendorff-perfused rat hearts using a potentiometric dye (di-8-ANEPPS).
Removal of the heart from anesthetized rats must be performed rapidly to avoid myocardial ischemia. If ischemia or insufficient coronary perfusion occurs, the heart will likely develop arrhythmias and may become infarcted. Additionally, these hearts will show insufficient fluorescence emission for informative recordings and subsequent analyses. Before loading the dye, myocardial cells need to be adequately perfused with Krebs-Henseleit solution to establish and maintain a physiologic electrolyte milieu for the stability …
The authors have nothing to disclose.
This work is supported by research grants from the National Institutes of Health (HL068915; HL088206) and contributions to the Cardiac Conduction Fund at Children’s Hospital Boston.
Material Name | Type | Company | Catalogue Number | Comment |
---|---|---|---|---|
CardioCMOS-SM128f | Equipment | RedShirt Imaging, Decatur, GA 30030 USA | ||
CardioPlex Software | Equipment | RedShirt Imaging, Decatur, GA 30030 USA | ||
LUXEON LED Light Source 460-490 nm | Equipment | Lumileds Lighting, US, LLC, San Jose, CA 95131 USA | LXHL-PB02 | |
ECG Amplifier Type 689 Hugo Sachs Elektronik | Equipment | Harvard Apparatus, Holliston, MA 01746 USA | 730149 | |
Dichroic Mirror 505 nm | Equipment | Semrock, Rochester, NY 14624 USA | FF505-SDi01-25×36 | |
Emission Filter 605 nm Long Pass | Equipment | SciMedia, Costa Mesa, CA 92626 USA | ||
THT Sideways | Equipment | SciMedia, Costa Mesa, CA 92626 USA | 25 BM-8 | |
Mini Ball Joint Holder | Equipment | Harvard Apparatus, Holliston, MA 01746 USA | BS4 73-0177 | |
Small Stimulation Electrode Set | Equipment | Harvard Apparatus, Holliston, MA 01746 USA | BS4 73-0160 | |
BM-6 Benchtop Vibration Isolation Platform | Equipment | Technology Inc., Inglewood, CA 90301 | 25 BM-6 | |
Monopolar ECG Electrode | Equipment | Harvard Apparatus, Holliston, MA 01746 USA | BS4 73-0200 | |
Roller Pump SCI 400 | Equipment | Watson-Marlow Bredel Inc., Wilmington, MA 01887 USA | 401U/D1 | |
Roller Pump MasterFlex Easy Load II | Equipment | Cole Parmer, Vernon Hills, Illinois 60061 USA | Model 77201-60 | |
Tubing Marprene #14 | Equipment | Watson-Marlow Bredel Inc., Wilmington, MA 01887 USA | 902.0016.016 | |
MasterFlex Tubing | Equipment | PharMed, Westlake, OH 44145 USA | 06485-25 | |
S48 Square Pulse Stimulator | Equipment | Grass Technologies, West Warwick, RI 02893 USA | Model S48 | |
SIU5 RF TRANSFORMER ISOLATION UNIT | Equipment | Grass Technologies, West Warwick, RI 02893 USA | Model SIU5 | |
5 Liter Water Jacketed Reservoir | Equipment | Radnoti Glass Technology Inc., Monrovia CA 91016 USA | 120142-5 | |
2 Liter Water Jacketed Reservoir | Equipment | Radnoti Glass Technology Inc., Monrovia CA 91016 USA | 120142-2 | |
0.5 Liter Water Jacketed Reservoir | Equipment | Radnoti Glass Technology Inc., Monrovia CA 91016 USA | 120142-0 | |
0.25 Liter Water Jacketed Reservoir | Equipment | Radnoti Glass Technology Inc., Monrovia CA 91016 USA | 120142-025 | |
10 ml Heating Coil | Equipment | Radnoti Glass Technology Inc., Monrovia CA 91016 USA | 158822 | |
Compliance Bubble Trap | Equipment | Radnoti Glass Technology Inc., Monrovia CA 91016 USA | 130149 | |
Luer Disconnect Cannula | Equipment | Harvard Apparatus, Holliston, MA 01746 USA | 72-1444 | |
3-Way stopcock, FLL to MLT, No Port Covers | Equipment | Harvard Apparatus, Holliston, MA 01746 USA | BS4 72-2630 | |
Thermocouple Thermometer | Equipment | Cole Parmer, Vernon Hills, Illinois 60061 USA | WU-91100-40 | |
Ultra Fine IT-Series Flexible Microprobe | Equipment | PhysiTemp Instruments Inc., Clifton, NJ 07013 USA | IT-24P | |
Oscilloscope Tektronix TDS 1002 | Equipment | Tektronix Inc., Beaverton, OR 97005 USA | TDS 1002B | |
2,3-Butanedione monoxime | Reagent | Sigma, St. Louis, MO 63132 USA | B0753 | |
Ketamine HCl | Reagent | Hospira Inc., Lake Forest, IL 60045 USA | RL-0065 | |
Xylazine | Reagent | Lloyd Inc., Iowa 51601 USA | LB15705A | |
E-TOXA-CLEAN® | Reagent | Sigma, St. Louis, MO 63132 USA | E9029 | |
Di-8-ANEPPS | Reagent | Invitrogen, Carlsbad, CA 92008 USA | D-3167 |