DESIGN & CONSTRUCT PELTIER DRIVEN COOLING & HEATING STAGE FOR LIGHT MICROSCOPE

(由美国国立卫生研究院/美国国立卫生研究院/P41 RR 01219拨款支持)。控制 整个实验过程中活体样本的温度，同时 通过视频延时显微镜观看，是维护的基础 他们的生存能力。此外，在许多情况下，试验性协议 要求细胞快速冷却，然后在细胞处于 通过高分辨率视频光学显微镜观察。例如，在 某些哺乳动物细胞内的所有微管都可以 通过快速冷却到6摄氏度来拆解，然后重新升温，然后允许 微管从中心体重新生长。在某些情况下，它是 希望能够解聚特定的微管 细胞，然后在观察细胞时用激光摧毁它的中心体 显微手术或通过光学捕捉来操纵它。为此，我们 设计并建造了一种廉价的帕尔蒂埃驱动的冷却系统 和加热显微镜阶段。这个阶段可以冷却里面的标本 玫瑰或灌注室(即使在浸油的情况下也是如此 目标)在不到6分钟内达到~4xC，保持不变 根据需要持续温度(&gt；60分钟)，然后在需要时 将标本加热至恒温。帕尔蒂埃设备 允许快速冷却/加热，因为大(T 在装置和样本之间产生。这一特点， 当与热传递函数的可逆性和 Peltier设备本身的低成本(&lt；每台20.00美元)使这成为一种 理想的温度控制阶段的实验在一个阶段 要求快速、可控、可持续的光学显微镜 温度会发生变化。Rieder，C.L.和R.W.Cole.1998年。灌流 用于高分辨率视频光学显微镜研究的小室 脊椎动物细胞单层：一些考虑和设计。在……里面 《视频显微镜》，G.Sluder和D.E.Wolf编辑。单元格中的方法 生物学56：253-275。

(Supported by the NIH/NCRR/P41 RR 01219 grant). Controlling the temperature of living specimens throughout an experiment, and while viewing by video time-lapse microcopy, is fundamental to maintaining their viability. In addition, in many cases an experimental protocol calls for rapidly cooling and then re-warming a cell while it is being viewed by high resolution video-light microscopy. For example, in certain mammalian cells all microtubules within the cell can be disassembled by rapidly cooling it to 6oC, and re-warming then allows the microtubules to re-grow from the centrosome. In some cases it is desirable to be able to de-polymerizes the microtubules in a specific cell, and then while viewing the cell destroy its centrosome by laser microsurgery or manipulate it by optical trapping. To this end we have designed and constructed an inexpensive Peltier-driven cooling and heating microscope stage. The stage can cool the specimen within a Rose or perfusion chamber (even when viewed with oil immersion objectives) to ~ 4xC in less than 6 minutes, hold it at this temperature for as long as needed (>60 min), and then when desired warm the specimen to homeostatic temperature. The Peltier device allows for the rapid cooling/heating due to the large (T that can be generated between the device and the specimen. This characteristic, when coupled to the reversibility of heat transfer function and the low cost of the Peltier devices themselves (<$20.00 ea.) makes this an ideal temperature control stage for experiments on the stage of a light microscope that call for rapid, controlled and sustained temperature shifts. Rieder, C. L. and R.W. Cole. 1998. Perfusion chambers for high-resolution video-light microscopic studies of vertebrate cell monolayers: some considerations and a design. In "Video Microscopy", G. Sluder and D. E. Wolf, Eds. Methods in Cell Biology 56:253-275.