Login G R, Dvorak A M
Am J Pathol. 1985 Aug;120(2):230-43.
We have demonstrated that microwave energy (MW) can be used in conjunction with chemical cross-linking agents in order to rapidly fix cell suspensions and tissue blocks for electron microscopy in 7-9 seconds. The optimal MW fixation method involved immersing tissues up to 1 cu cm in dilute aldehyde fixation and immediately irradiating the specimens in a conventional microwave oven for 9 seconds to 50 C. Ultrastructural preservation of samples irradiated by MW energy was comparable to that of the control samples immersed in aldehyde fixative for 2 hours at 25 C. Stereologic analysis showed that tissue blocks fixed by the MW fixation method did not cause organelles such as liver mitochondria and salivary gland granules to shrink or to swell. Potential applications for this new fixation technology include the investigation of rapid intracellular processes (eg, vesicular transport) and preservation of proteins that are difficult to demonstrate with routine fixation methods (eg, antigens and enzymes).
我们已经证明,微波能量(MW)可与化学交联剂结合使用,以便在7至9秒内快速固定细胞悬液和组织块用于电子显微镜检查。最佳的微波固定方法是将体积达1立方厘米的组织浸入稀醛固定液中,然后立即在传统微波炉中于50℃下照射标本9秒。经微波能量照射的样品的超微结构保存情况与在25℃下浸入醛固定剂2小时的对照样品相当。体视学分析表明,采用微波固定方法固定的组织块不会导致肝线粒体和唾液腺颗粒等细胞器缩小或肿胀。这项新固定技术的潜在应用包括对快速细胞内过程(如囊泡运输)的研究以及对常规固定方法难以显示的蛋白质(如抗原和酶)的保存。