Valentino K L, Crumrine D A, Reichardt L F
J Histochem Cytochem. 1985 Sep;33(9):969-73. doi: 10.1177/33.9.2991364.
We present methods for embedding brain tissue in Lowicryl K4M embedding medium and localizing antigens using postembedding immunogold techniques. After perfusion fixation with 4% paraformaldehyde and 0.1% glutaraldehyde in 0.1 M cacodylate buffer, blocks of rat brain were placed in 2% aqueous uranyl acetate for 1 hour, dehydrated in 50%, 70%, and 95% ethanol, infiltrated with Lowicryl/ethanol mixtures (1:2 for 10 min, 1:1 for 15 min) and 100% Lowicryl (20 min and 25 min). Polymerization was carried out under UV light for 24-48 hours at room temperature. Several neural antigens, including three different synaptic vesicle proteins and an enzyme associated with the postsynaptic density, were localized by this technique, indicating that this procedure may have wide applicability.
我们介绍了将脑组织嵌入Lowicryl K4M包埋介质以及使用包埋后免疫金技术定位抗原的方法。在用0.1M二甲胂酸钠缓冲液中的4%多聚甲醛和0.1%戊二醛进行灌注固定后，将大鼠脑块置于2%乙酸铀水溶液中1小时，依次在50%、70%和95%乙醇中脱水，用Lowicryl/乙醇混合物（1:2比例10分钟、1:1比例15分钟）和100% Lowicryl（20分钟和25分钟）进行渗透。在室温下紫外光下聚合24 - 48小时。通过该技术定位了几种神经抗原，包括三种不同的突触囊泡蛋白和一种与突触后致密物相关的酶，表明该方法可能具有广泛的适用性。
