McGrath C M, Grudzien J L, Decker D A, Robbins T O
Grace Bio-Oncology Laboratory, Inc., Pontiac, MI 48342.
Biotechniques. 1991 Sep;11(3):352-4, 356, 358-61.
A new method (Freeze-Transfer) is described for performing high-resolution immunocytochemistry for soluble cell proteins on frozen sections of biological tissues that involves thaw-mounting frozen tissue sections directly onto the surface of nitrocellulose thin films instead of directly onto glass slides. This technically straight-forward change in methodology resulted in chromogenic immunocytochemical assays for Her-2 and EGF receptors that were 1-2 orders of magnitude more sensitive while still fully utilizing the diagnostic resolving power of light microscopy. The effects of membrane pore size and surface chemistry on the resolution and intensity of Her-2 signal suggest that the enhanced sensitivity of Freeze-Transfer was caused by the cytologically coherent transfer of target molecules normally lost from cut surfaces of cells mounted on nonporous glass during assay.
本文描述了一种新方法（冷冻转移法），用于在生物组织的冷冻切片上对可溶性细胞蛋白进行高分辨率免疫细胞化学分析，该方法将冷冻组织切片直接解冻固定在硝酸纤维素薄膜表面，而不是直接固定在载玻片上。这种技术上简单的方法改变，使得针对Her-2和表皮生长因子（EGF）受体的显色免疫细胞化学检测灵敏度提高了1 - 2个数量级，同时仍充分利用了光学显微镜的诊断分辨能力。膜孔径和表面化学对Her-2信号分辨率和强度的影响表明，冷冻转移法灵敏度提高的原因是在检测过程中，通常从固定在无孔玻璃上的细胞切面丢失的靶分子在细胞学上实现了连贯转移。
