Simple Myocyte/Langendorff #120108EZ
For researchers requiring a simple, easy to use and inexpensive isolated heart system for Langendorff experiments or myocyte isolation. The system may be configured for a gravity-fed constant pressure mode, or for constant flow, the later with a user supplied peristaltic pump. The system comes complete with a two bar lab stand and stabilizer bar, water jacketed buffer reservoir and a heating coil with a built in bubble trap. In the constant flow mode, a user supplied peristaltic pump draws buffer from the reservoir, passes it through the heating coil, where the bubble trap removes unwanted air bubbles and then passes down to the aortic cannula (one 2.4mm glass cannula, and one stainless steel cannula with luer locking hub are provided). A 100ml water jacketed open top chamber serves to collect the effluent and provide some warming for the exposed heart. For myocyte isolations, solutions may be recirculated via a user provided peristaltic pump. Ideal for student experiments or basic myocyte isolation requirements in the research lab.
Radnoti High Yield Myocyte Isolation System #120107EZ
Can be used for Constant Pressure or Constant Flow Langendorff in both recirculating and non recirculating models.
- stand assembly
- water jacketed heart chamber with a collecting trap
- perfusate manifolds and valves
- 3 water jacketed reservoirs with aerators
- 3 water jacketed bubble traps
- water jacketed perfusate lines
- peristaltic pump
- thermal circulator pump.
Overview of Myocyte Isolation
Isolation of viable myocytes from perfused hearts is the initial and most critical step in many optical and/or electro physiological experiments or for long term culturing. The system can be used for either constant pressure or constant flow Langendorff preparations. If the experimenter decides to use the more common constant flow isolation , a manifold is used to select the appropriate solution that is then pumped through a bubble trap and into the heart. First, crystalloid solutions (Kreb’s or Tyrode’s) are used to remove blood cells and stabilize the preparation, then a solution containing EGTA is used to remove residual calcium. A solution containing collagenase and sometimes other digestive enzymes is then recirculated through the preparation until the heart becomes flaccid, indicating that the intercellular matrix holding the cells together is dissolving. The partially digested tissue is then cut into small chunks and placed in a shaking bath and subjected to continued dissociation treatment until individual myocytes are released. The protocol may require several changes of solution to select the myocyte fraction that has the best yield and viability, the latter determined through the use of microscopic examination in the presence of trypan blue or other vital dye staining. To increase viability, step changes in the calcium concentration of the incubation media are used to return the myocyte to a normal ionic environment prior to their use. If the experimenter decides to use a constant pressure isolation, the reservoir(s) are elevated to the appropriate hydrostatic head and a manifold directs the selected solution to the appropriate bubble trap and then to the heart. A valve at the bottom of the heart chamber diverts the perfusate either to waste or to the pump for recirculating back to the source of the solution.