免疫酶双重染色中抗体洗脱的进一步研究

本实验进一步检查了三种常用的洗脱方法从切片上除去免疫酶组织化学染色后的抗体的效果。结果表明,PAP法染色后,氧化法的抗体洗脱完全,效果可靠;酸洗法和酸洗/二甲基甲酰胺法的效果视不同抗体而不同,二甲基甲酰胺单用或用于酸洗后似无效。所用方法均不能从ABC法染色的垂体组织切片上完全除去抗体复合物。因之,将ABC法用于第一抗体来源于同一动物种的双重染色中,来显示第一种抗原时,要特别注意排除假性双标记。     

葡萄糖氧化酶—二氨基联苯胺—硫酸镍铵法在免疫酶双重染色技术中应用

作者在豚鼠胰腺组织石蜡切片的PAP免疫酶双重标记染色中,分别使用葡萄糖氧化酶-二氨基联苯胺-硫酸镍铵(Glucose oxidase-DAB-Nickel,GDN)和单纯DAB显示生长抑素(Somatostatin,SOM)及5-羟色胺(5-hpdroxytryptamin,5-HT)免疫反应细胞,获得良好的染色结果。该方法步骤简单,结果明确,背景清楚,是取得理想的免疫酶双重染色结果的新途径。     

DNA染色与免疫组织化学双重染色技术

随着计算机技术的普及和图象定量分析方法的发展 ,病理学已经开始应用计算机图象分析技术来定量研究有关病变形态结构特点 ,探讨其在诊断、分型、分类、预后判断中的应用等问题。我们采用Feulgen Rossenbeck改良DNA染色法[1] 和免疫组织化学EnVision法结合的双重染色方法 ,在石蜡切片中同时显示乳腺癌细胞DNA和癌基因CerbB 2的蛋白表达产物 ,该法对应用病理学图象分析技术研究肿瘤细胞DNA及癌基因蛋白的表达及其两者关系提供了一种简便、准确的方法。1　材料和方法1 1　材料 :取自本科室乳腺癌标本 ,标本经10 %中性福尔马林固定 ,常规…     

微波在抗酸染色中的应用改进

抗酸染色以Ziehe—Neelsen抗酸杆菌法的应用最为广泛,但操作过程中温度及复染背景难以控制,在总结了我科10例麻风活检标本抗酸染色基础上,我们对抗酸染色法进行改良,效果显著,现介绍如下。     

微波在Cajal染色中的应用

在Cajal氨酒精去神经组织块镀银染色过程中,对固定、浸银、还原等过程进行微波辐射处理,结果表明微波辐射能明显促进固定、浸银、还原作用,使组织块镀银染色时程大大缩短。     

微波技术在骨骼肌染色中的应用

均高于对照组;在形态学的实验教学中,骨骼肌纤维的铁苏木精染色是观察骨骼肌横纹的经典方法,传统的铁苏木精染色方法存在着耗时较长且染色不够均匀等不足。近几十年来,微波照射（Microwave irradiation,MWI）被成功地引用到组织固定、组织化学、免疫组织化学等研究中。MWI具有波长短、频率高,能使微波场内某些化学物质迅速地穿透物体,促进液体浸入标本等特点。本实验将微波辐射应用于骨骼肌纤维染色,并与常规室温下的染色进行对照,经反复探索得到较为满意的效果。     

特殊染色及免疫组化双重染色在恒河猴糖尿病模型中的应用

目的在链脲佐菌素（STZ）诱导恒河猴糖尿病动物模型基础上,用特殊染色、免疫组化双染方法显示胰腺等组织中的特征性病变。方法健康恒河猴5只,小剂量（30mg／kg）多次静脉注射STZ,濒死状态时将动物安乐死。取胰腺、心脏、肾脏、脾脏、肝脏、眼球、脑等器官制成石蜡切片,用HE、PAS、Masson、天狼星红和甲苯胺蓝等方法进行特殊染色,用免疫组化双重染色法同时显示胰岛A、B细胞。结果模型动物的胰岛萎缩,数量减少。Masson染色见外分泌部间质内纤维增生。局灶性慢性肾炎,病变区肾间质纤维组织增生,部分肾小球及肾小管萎缩。天狼星红染色见脾脏中央动脉管壁增厚。免疫组化双染见胰岛A胰高血糖素表达增多,胞浆呈棕褐色。B胰岛素表达减少,胞浆粉红色。结论HE染色结合特殊染色和免疫组化双重染色可较好地对STZ诱导糖尿病动物模型进行组织学评价。     

微波在蛋白质电泳染色和脱色中的应用

经过实验摸索和实践 ,我们发现一种利用微波处理在10ｍｉｎ内可使聚丙烯酰胺凝胶染色及脱色完成的方法 .ａ 将电泳后的凝胶 (厚度 0 75ｍｍ)取出 ,置于培养皿中用蒸馏水冲去残留缓冲液 .ｂ 加入染色液将凝胶完全浸泡其中 ,放入微波炉内高火档照射 2 0ｓ,停留 1ｍｉｎ ,再照射 10ｓ.ｃ 取出染色液 (回收还可再用 2次 ) ,用蒸馏水冲去凝胶上残留染色液 .ｄ 加入脱色液 ,高火档照射 2 0ｓ ,取出脱色液 ,然后加入新鲜脱色液再照射 2 0ｓ,重复5～ 6次 ,直至背景清晰透明 .实践中我们认为应注意以下几点 :ａ 盛试剂的培养皿上面应盖一稍大的培养…     

骨髓标本中免疫组织化学及DNA原位末端标记双重染色技术的应用

为扩展免疫酶标记分子病理学技术在血液病基础与临床研究中的应用,我们探索了碱性磷酸酶抗碱性磷酸酶（A-PAAP）免疫组化和DNA原位末端标记（ISEL）技术在骨髓涂片和切片（乙二醇甲基丙烯酸酯GMA冷包埋）中联合应用（双重染色）的方法,结果令人满意,已初步用于白血病分化抗原及细胞凋亡的共同检测。     

微波快速免疫荧光组化染色方法的研究

本文应用微波辐射方法加速免疫荧光组化染色(间接和直接法),分别定位15种不同组织抗原,并应用连续切片同时用两种不同的孵育方法即微波辐射和常规孵育方法进行比较。结果证明,经微波辐射后免疫荧光组化染色时间大大缩短,背景染色明显好于常规法,阳性率和阳性强度与常规法基本一致。     

抗人肺腺癌细胞株SPC-A-1的单克隆抗体的制备和鉴定

人肺腺癌细胞株SPC-A-1,经抗人正常肺抗血清复被后免疫小鼠,脾细胞和同系NS-1或SP 2/0融合,获得17株单克隆抗体。本文报道其建株过程及其中7株单克隆抗体的一些特征。LAC-122、LAC-163与LAC-210对人体肺腺癌和肺鳞癌的阳性反应率分别为75%、78.3%和75%,除LAC-122对一例胃癌细胞株有微弱交叉反应外,三株单抗对肺小细胞癌, 其它肿瘤组织及正常、胚胎组织均呈阴性。对利用抗体复被瘤细胞及肺癌相关抗原问题作了较详细的讨论.     

A method for localizing embryonal laticifers by combined conventional and fluorescence microscopy

Summary The authors describe a simple method based on malachite green and acid fuchsin for the detection of laticifers during the embryogenesis of someEuphorbiaceae plants by conventional and fluorescence microscopy. The strong sensitivity and specificity of the method make it suitable for the ontogenetic studies of laticifers. The results obtained are discussed in the context of the reactive mechanism of the staining and of the chemical composition of the embryonal laticifers.This paper was presented in part at the International Meeting on Botanical Microscopy, organized by The Royal Microscopical Society in York, July 9–13, 1979.     

Fast and efficient egg collection and antibody staining from large numbers of Drosophila strains

 The genetic dissection of any developmental processes requires mutagenesis protocols and the subsequent phenotypic screen of the established mutant strains. Whereas external structures such as the Drosophila cuticle are relatively easy to score without the need of further manipulations, the analyses of internal structures such as the nervous system often requires the use of antibodies to detect abnormalities. Here we describe an improved method to: (a) simultaneously collect Drosophila eggs from large number of fly strains, (b) process them fast for antibody staining and (c) facilitate rapid subsequent screening. Received: 5 February 1997 / Accepted: 23 March 1997     

免疫酶双重染色法显示再生神经中的神经生长因子与睫状节神经营养因子

本研究以成人正中神经切割伤后２～３个月的神经干为材料,冰冻切片,用免疫双重染色技术显示了神经生长因子与睫状节神经营养（诱向）因子在再生的周围神经组织中的表达与分布。神经生长因子选用ＡＰＡＡＰ法．其阳性产物呈红色;睫状节神经营养（诱向）因子选用ＡＢＣ系统,４氯－１－萘酚显色,阳性产物为褐色。光镜下观察：神经生长因子的阳性反应产物出现在正中神经切割伤后再生的神经纤维中,高倍镜下可见其阳性产物分布在轴索,而在雪旺氏细胞中没能见到呈红色的阳性反应产物;睫状节神经营养（诱向）因子分布在一些细胞体积大、核大呈增生活跃状态的雪旺氏细胞中。红与褐双色反应产物色调清晰,效果较好。研究结果提示：睫状节神经营养（诱向）因子与神经生长因子在人周围神经再生过程中起着十分重要的作用。     

神经细胞尼氏体和神经髓鞘的双重组合染色法与应用

探讨了显示实验动物脊髓组织中的神经尼氏体和神经髓鞘等组织成分的双重组合染色法,通过分别选用孔雀石绿(Malachite green)和橙黄G、磷钨酸(Orange G,Phosphotungstic acid)组合染色(简称MG-OP法)。已能够显示狗脊髓神经尼氏体呈绿色,细胞核呈黄色。神经纤维髓鞘轴突呈黄色,神经膜和结缔组织纤维呈绿色,背景呈淡黄色。MG-OP法克服了原法成分单一,色彩效果差,所建立的双重组合染色法,对比清晰,色彩鲜艳,方法简便的较好染色方法。     

Immunoenzyme method for differential assay of free and bound ABA

We performed the immunochemical study of two solid-phase competitive ELISA systems differing in their specificity toward free and bound ABA. A possible application of these systems for the quantification of natural ABA forms without their preliminary separation and purification in a single sample of plant material was demonstrated.     

Identification of alpha-1-proteinase inhibitor-containing cells in pancreatic islets

Summary By a regular immunoperoxidase method, alpha-1-proteinase inhibitor (Api) was demonstrated in pancreatic islets in individuals with and without genetic deficiency of Api. Subsequently a double immunoperoxidase method, with two different chromogens (diaminobenzidine-brown and 4-chloro-1-naphthol-blue), was applied on the same tissue section in order to identify cells containing Api and cells secreting polypeptide hormones. Api-positive cells and hormone-secreting cells were found to be mutually exclusive indicating that Api is synthesized by previously unrecognized islet cell. The population of Api cells was significantly higher in persons with genetic deficiency than in other individuals, implying a possible compensatory hyperplasia of those cells triggered by a low level of circulating Api.     

Immunocytochemical demonstration of polyamines in nucleoli and nuclei

Although biochemical studies have shown that polyamines (PAs) occur in the nucleus, only few studies have examined the intranuclear distribution of these organic cations. By immunocytochemistry, we have previously demonstrated that PAs are located in ribosomes. We now show that PAs also are present in both nucleoli and nuclei of a variety of cell types. Detection of nucleolar and nuclear PAs required novel pretreatment procedures involving protease and/or DNase digestion of specimens prior to immunoreaction. Double fluorescence staining confirmed the localizations. This suggests that PAs may be important to the formation of ribosomes in nucleoli, as well as adds support to biochemical studies suggesting that PAs are involved in many biological events in the nucleus. Further biochemical studies will be needed to substantiate this hypothesis.