New York State Psychiatric Institute and Columbia University, Unit 42, 1051 Riverside Drive, New York, NY 10032, USA; Macedonian Academy of Sciences and Arts (MASA), Bul. Krste Petkov Misirkov 2, Skopje 1000, Macedonia; School of Medicine, University Ss. Cyril & Methodius, Vodnjanska 17, Skopje 1000, Macedonia.
Institute of Chemistry, Faculty of Natural Sciences and Mathematics, University Ss. Cyril & Methodius, Arhimedova 5, Skopje 1000, Macedonia.
J Neurosci Methods. 2014 Jun 15;230:20-9. doi: 10.1016/j.jneumeth.2014.04.006. Epub 2014 Apr 18.
Golgi stains are notoriously capricious, particularly when applied to human brain. The well-known difficulties, which include complete failure of impregnation, patchy staining, unstable staining, and extensive crystalline deposits in superficial sections, have discouraged many from attempting to use these techniques. A reliable method that produces uniform impregnation in tissue from human autopsies and experimental animals is needed.
The method described, "NeoGolgi", modifies previous Golgi-Cox protocols (Glaser and Van der Loos, 1981). Changes include: much longer time (>10 weeks) in Golgi solution, agitation on a slowly rocking platform, more gradual infiltration with Parlodion, more thorough removal of excess staining solution during embedding, and shorter exposure to ammonia after infiltration.
The procedure has successfully stained over 220 consecutive frontal or hippocampal blocks from more than 175 consecutive human autopsy cases. Dendritic spines are easily recognized, and background is clear, allowing examination of very thick (200 μm) sections. Stained neurons are evenly distributed within cortical regions. The stain is stable for at least eight years. Most importantly, all stained neurons are apparently well-impregnated, eliminating ambiguity between pathology and poor impregnation that is inherent to other methods.
Most methods of Golgi staining are poorly predictable. They often fail completely, staining is patchy, and abnormal morphology is often indistinguishable from poor impregnation. "NeoGolgi" overcomes these problems.
Starting with unfixed tissue, it is possible to obtain Golgi staining of predictably high quality in brains from human autopsies and experimental animals.
高尔基染色法非常不稳定，尤其是在应用于人脑时。众所周知，这种技术存在许多困难，包括完全浸渍失败、染色不均匀、染色不稳定以及在浅层切片中出现大量结晶沉积物等问题，这使得许多人望而却步，不愿尝试使用这些技术。目前需要一种可靠的方法，能够在人体和实验动物的组织中产生均匀的浸渍效果。
本文描述的方法“NeoGolgi”对以前的高尔基-考克斯（Golgi-Cox）方案进行了修改（Glaser 和 Van der Loos，1981）。更改包括：在高尔基溶液中浸泡时间更长（>10 周）、在缓慢晃动的平台上搅拌、用 Parlodion 更渐进地渗透、在包埋过程中更彻底地去除多余的染色溶液以及在渗透后更短时间暴露于氨水中。
该程序已成功对 175 例连续尸检案例中的 220 个连续额或海马体块进行了染色。树突棘很容易识别，背景清晰，允许检查非常厚（200μm）的切片。染色神经元在皮质区域内均匀分布。该染色稳定至少八年。最重要的是，所有染色的神经元显然都被很好地浸渍，消除了其他方法中固有的病理学和浸渍不良之间的歧义。
大多数高尔基染色方法都难以预测。它们经常完全失败，染色不均匀，异常形态学通常与浸渍不良难以区分。“NeoGolgi”克服了这些问题。
从未固定的组织开始，有可能在人体和实验动物的大脑中获得可预测的高质量高尔基染色。
