Shellman Yiqun G, Ribble Deborah, Yi Ming, Pacheco Theresa, Hensley Mark, Finch Dudley, Kreith Frank, Mahajan Roop L, Norris David A
University of Colorado Health Science Center, Denver, CO, USA.
Biotechniques. 2004 Jun;36(6):968-72, 974-6. doi: 10.2144/04366ST01.
Hyperthermia, the procedure of exposing cells to a temperature between 42 degrees and 49 degrees C, has been shown to be a promising approach for cancer treatment. To understand the underlying mechanisms of hyperthermic killing of cancer cells, it is critical to have an accurate temperature measurement technique and a heating method with high reproducibility. To this end, we have developed a method using fine thermocouples with fast response time to measure the temperatures in multiple wells of a 96-well plate. The accuracy of temperature measurement was +/- 0.2 degree C. Such a capability allows a complete record of the time and temperature of the treatment procedure and helps define an accurate thermal dose. We have also compared several methods for heating 96-well plates and found that use of copper blocks in contact with the lower surface of the 96-well plate in an incubator provides a highly reproducible heating method. The common method of using water bath to heat cells in vitro resulted in a decrease of cell viability even at the control temperature of 37 degrees C and a decrease in the reproducibility of certain biological assays. In summary, using these improved techniques, proposed thermal dose can be defined more precisely, and highly reproducible heating in vitro can be achieved.
热疗，即将细胞暴露于42摄氏度至49摄氏度之间的温度的过程，已被证明是一种很有前景的癌症治疗方法。为了理解热疗杀死癌细胞的潜在机制，拥有精确的温度测量技术和高重现性的加热方法至关重要。为此，我们开发了一种使用响应时间快的精细热电偶来测量96孔板多个孔中温度的方法。温度测量的精度为±0.2摄氏度。这种能力可以完整记录治疗过程的时间和温度，并有助于定义精确的热剂量。我们还比较了几种加热96孔板的方法，发现使用铜块与培养箱中96孔板的下表面接触可提供高度可重现的加热方法。在体外使用水浴加热细胞的常用方法即使在37摄氏度的对照温度下也会导致细胞活力下降，并降低某些生物学测定的重现性。总之，使用这些改进的技术，可以更精确地定义提议的热剂量，并在体外实现高度可重现的加热。
