Oates K, O'Connor P, Gardner D L, Orford C R
Ultrastruct Pathol. 1984;6(2-3):239-46. doi: 10.3109/01913128409018579.
A scanning electron microscopic technique for the examination of bulk, fresh, hydrated human tissue is described. Samples of fresh tissue are frozen in liquid nitrogen against a mirror-finished copper block and planed in a cryoultramicrotome before transfer to a low temperature scanning electron microscope. After sublimation of water from the specimen surface, the tissue is examined in secondary electron and backscattered electron modes. Adjacent pieces of tissue, and those retrieved after backscattered electron observation, can be readily prepared for and examined by light and by conventional transmission electron microscopy. The method has been tested with multiple blocks taken from 6 cases of human breast carcinoma. In the backscattered electron mode, the infiltrating columns of neoplastic cells can be distinguished from mammary adipose and fibrous tissue. Within a carcinoma, the collagenous stroma, carcinoma cells, and perivascular and perineural infiltrates can be identified. These features have been contrasted with those obtained by light microscopy, by low temperature scanning, and by transmission electron microscopy. This use of backscattered electron imaging for the investigation of unfixed hydrated tissue offers the possibility that the technique could be of considerable value in the microscopy of very small samples in which, because of a need for subsequent biochemical, histochemical, and immunologic investigation, fixation and dehydration are to be avoided.
本文描述了一种用于检查整块、新鲜、含水人体组织的扫描电子显微镜技术。将新鲜组织样本在液氮中对着镜面抛光的铜块冷冻，然后在冷冻超薄切片机中切片，再转移至低温扫描电子显微镜下。在标本表面的水升华后，以二次电子和背散射电子模式检查组织。相邻的组织块以及在背散射电子观察后取回的组织块，可以很容易地制备用于光镜和传统透射电子显微镜检查并进行观察。该方法已用取自6例人类乳腺癌的多个组织块进行了测试。在背散射电子模式下，可以将肿瘤细胞的浸润柱与乳腺脂肪和纤维组织区分开来。在癌组织内，可以识别出胶原基质、癌细胞以及血管周围和神经周围浸润。这些特征已与通过光学显微镜、低温扫描和透射电子显微镜获得的特征进行了对比。这种利用背散射电子成像来研究未固定的含水组织的方法表明，该技术在极少量样本的显微镜检查中可能具有相当大的价值，因为出于后续生化、组织化学和免疫学研究的需要，要避免固定和脱水。
