Cryostat for cutting flash-frozen tissue sections

所需的设备是徕卡低温恒温器，用于切割先前快速冷冻的组织的薄片。Gardiner由NSERC资助的研究的一个主要部分涉及确定蛋白质的水平，以及控制大鼠后肢肌肉α运动神经元的神经元中编码这些蛋白质的信使RNA的水平。另一个研究方向涉及测量肥胖和糖尿病对大鼠血液流动的影响及其在各种不同组织中的调节。在这两种情况下，从大鼠身上取出的组织必须迅速冷冻，然后在大约零下20摄氏度的温度下切片，以便随后处理这些组织，以可视化我们感兴趣的元素。Stecina由NSERC资助的研究项目涉及识别成年啮齿动物神经系统中的脊髓中间神经元及其功能网络。在几乎所有这些实验中，除了电生理学研究外，还需要在事后验证刺激电极在大脑中的放置。此外，对于神经元间网络的电生理特征，神经解剖学方法，如活动依赖标记，脊髓记录位置的识别也在每一次执行的实验后实际使用。此外，对每次电生理实验后采集的脊髓和脑组织进行薄片切割，需要经常使用低温恒温器(每周至少1-2天)。对于考利来说，NSERC资助的步进和平衡神经基础研究需要使用低温恒温器进行损伤验证，涉及中间神经元和运动神经元的逆行标记研究，以及与神经递质受体分布相关的免疫组织化学研究。我们目前使用的低温恒温器已有15年的历史，使用量很大，而且正在损坏。购买所需的设备将使所有这些研究计划不受干扰地继续进行，这些研究计划对我们理解如何运动、参与运动的神经网络如何由高等中枢控制以及环境如何影响神经和其他组织的基因表达和功能具有影响。当目前使用的低温恒温器停止正常工作时，我们预计随时会发生这种情况，因为它的老化、更换部件的不可用以及目前的大量使用，三名研究人员的研究计划将严重受损，以至于他们各自的研究计划的全部部分将无法完成。

The equipment requested is a Leica cryostat for cutting of thin sections of previously rapidly-frozen tissue. A major part of Gardiner's NSERC-funded research involves determining the level of proteins, and of the messenger RNAs which code for these proteins, in the neurons in rats which control the hindlimb muscles, the alpha-motoneurons. Another research direction involves measuring the effects of obesity and diabetes in rats on blood flow and its regulation in a variety of different tissues. In both cases, tissues removed from the rat must be rapidly frozen, and later sectioned at a temperature of about minus 20 degrees C, to treat the tissues subsequently to visualize the elements we are interested in. Stecina's NSERC-funded research program involves the identification of spinal interneurons and their functional networks in the adult rodent nervous system. In almost all of those experiments, in addition to the electrophysiological studies, verification of the placement of stimulation electrodes in the brain is required post-hoc. Also, for the electrophysiological characterization of interneuronal networks, neuroanatomical methods, such as activity dependent labelling, identification of recording sites in the spinal cord are also used practically after each of the performed experiment. In addition, the cutting of thin sections from spinal cord and brain tissue harvested after each electrophysiological experiment warrants the use of a cryostat on a frequent basis (minimum 1 - 2 days per week). For Cowley, NSERC-funded investigations of the neural basis of stepping and balance require the use of a cryostat for lesion verification, studies involving retrograde labeling of interneurons and motoneurons, as well as for immunohistochemistry related to the neurotransmitter receptor distribution. We currently use a cryostat that is 15 years old, which is heavily used, and is breaking down. The purchase of the requested equipment will allow all of these research programs, which have implications for our understanding of how we move, how the neural networks involved in movements are controlled by higher centres and how the environment influences the gene expression and function of nervous and other tissues, to continue without interruption. When the currently-used cryostat ceases to function properly, which we expect will occur at any time due to its age, the unavailability of replacement parts, and current heavy usage, the research programs of the three investigators will be severely compromised, such that entire portions of their respective research programs will be impossible to complete.