Myocyte Isolation Systems
对于光学影像及电生理实验，从灌注心脏分离心肌细胞是最初的非常重要的一步。灌流系统可以进行恒压或者横流langendorff灌流。加入实验者决定通常使用横流灌流分离，一个分流管可以用来选择合适的溶液，通过蠕动泵灌入心脏。首先，Kreb's溶液用来清洗血细胞，冰鞋稳定标本。接下来。含有EGTA的溶液用来去除残留的钙。在通过一种含有胶原酶或者消化酶的溶液循环灌注标本，知道心脏变得松软，表明细胞间保持排列的物质消融了。将部分消化的组织小块放入摇床水浴，连续进行强制的溶解处理，直到释放出心肌细胞。整个实验过程，需要切换几次溶液来选择最容易活的心肌细胞。为了增加生命力，需要阶段性改变孵育介质的钙浓度，在使用前使心肌细胞返回到它们适应的离子环境中。假如实验者决定用恒压灌流模式隔离，灌流液瓶需要被高架,分流管直接选择溶液，将溶液经气液闸灌入心脏。心脏腔体下面的阀门可选择奖灌注液排掉或通过泵进行再循环回源溶液。
有两个配置可以选择，简化的120107心肌细胞分离系统。复杂的120108心肌细胞分离系统
120107简化的心肌细胞分离系统
心肌细胞分离灌流系统
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.
Includes:
?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.
现货Myocyte Isolation Systems Simple Myocyte/Langendorff #120108EZ现货Myocyte Isolation Systems Simple Myocyte/Langendorff #120108EZ现货Myocyte Isolation Systems Simple Myocyte/Langendorff #120108EZ