快速石蜡切片在小块活检组织中的应用

目前,病理科普遍开展的常规石蜡技术,制片程序时间较长,三个工作日才能发出病理诊断报告;而有时在某些特殊情况下,送检组织已固定,不能行冰冻切片检查时,就必须在很短的时间内制好片。目     

全自动密闭式组织脱水机在石蜡切片组织处理中的应用

全自动密闭式组织脱水机问世以来,以其安全、高效和良好的处理效果等优点,深受病理学、解剖学、动物实验和组织胚胎等技术人员的青睐。我科自2002年使用全自动密闭式组织脱水机处理组织以来,组织的处理效果明显改观,石蜡切片质量显著提高,现将应用经验简介如下。材料和方法1·材料1·1标本组织来源于我院手术的活检标本。1·2仪器和设备樱花VIP-E150型全自动密闭式组织脱水机为日本原装进口,附带不锈钢脱水篮1只,可容150个包埋盒;塑料包埋盒为国产。1·3试剂冰乙酸,甲醛原液,95%乙醇,无水乙醇,二甲苯,石蜡(58-60℃)均为国产试剂。AAF固定…     

不同组织在石蜡包埋中的方法和体会

在石蜡切片制作过程中,组织包埋是一重要环节,不同组织和病变的包埋则有所不同,包埋位置直接影响切片和对病变的观察,现将我们在工作中的包埋方法和体会介绍如下：     

石蜡切片法中细长或薄片状材料的包埋

石蜡切片法制作细长或薄片状生物材料的横切面切片时,可进行下列简便而有效的石蜡包埋方法：先在包埋纸盒底部倒上少许熔蜡,形成约1mm厚的软蜡层;待软蜡层凝固前,再向纸盒中倒满熔蜡,然后迅速将渗好蜡的材料直立地放入熔蜡中,并将材料轻压在纸盒底部的软蜡层上。     

石蜡切片贴片法的改良

传统的石蜡切片贴片操作费时,而且效果不太理想。作者经过多次摸索试验,对其进行了改良,取得了良好的效果。     

快速石蜡切片法在免疫组化染色中的应用

免疫组化染色在病理诊断中发挥着重要的作用,较多的应用于肿瘤的诊断和鉴别诊断,而制作出良好的组织切片是免疫组化染色的基础和前提。快速石蜡制片是病理检验的常规技术之一,我们将常     

植物组织石蜡切片的扫描电镜观察方法研究

石蜡切片的扫描电镜观察法有其独到之处:集光镜和扫捕电镜特长于一体,在大量的石蜡切片光镜观察的基础上,挑选具有研究线索的切片,采用此法转移到扫描电镜下作高分辩研究,既可普查切片全貌,又可处得切片中亚微结构的三维图像,这对结构的准确分辩十分有利,且便于作连续切片观察。本文简要介绍这一实验技术。     

植物冰冻切片条件的优化及其与石蜡切片在组织化学应用中的比较

冰冻切片是植物组织学研究中一项重要的实验技术,冷冻温度和冷冻时间是决定切片质量的关键因素。通过比较15种冰冻切片条件,得出植物组织直接冰冻切片较适宜的冷箱温度、冷台温度和冷冻时间。同时,通过对5种植物的不同组织进行组织化学染色,比较了新鲜材料直接冰冻切片与常规石蜡切片在不同化学成分鉴定上的异同及各自的适用范围。结果表明,对于多糖、蛋白质和角质,两种切片方法的鉴定结果比较一致,但对于脂肪只能采用冰冻切片技术。研究结果对植物组织学实验和研究方法的改进具有一定的参考价值。     

动物组织石蜡切片H-E染色的快速方法

动物组织石蜡切片及染色技术是普通生物学及动物学实验中必需的实验技能.经过多年的积累和摸索,对组织切片中的苏木精-尹红( hematoxylin - eosin,H-E)染色技术进行了改进,取得了良好的教学效果和实验效果.     

兔卵巢组织石蜡切片技术的改良

针对常规石蜡切片方法步骤繁多,耗时长,所用的二甲苯试剂对人体健康有较大毒害性的缺陷,探讨提高制作兔卵巢组织石蜡切片质量的方法。试验从脱水和透明环节进行技术改良,用正丁醇与无水乙醇混合剂取代常规的脱水剂和透明剂。该混合剂具有良好的脱水和透明作用。实验结果表明：通过该技术改良,镜下观察组织结构清晰可辨、颜色鲜艳、对比度好、层次分明,组织切片效果良好。试验改良技术的工艺简单,具有质优、省时、快捷和实用性较好等优点,有推广应用价值。     

两种脱钙法植被鼠尾病理切片的比较

目的探讨制作大鼠尾巴标本石蜡切片的方法。方法采用盐酸脱钙液运用两种脱钙方法制作大鼠尾巴标本石蜡切片。结果两种方法制作的石蜡切片完整、无破碎,HE染色观察组织结构细胞形态完整,核浆分明,红蓝适度。结论两种方法都能制作理想大鼠尾巴标本石蜡切片,可保证病理诊断质量。     

Staining Paraffin Sections Without Prior Removal of the Wax

Paraffin sections are usually rehydrated before staining. It is possible to apply aqueous dye solutions without first removing the wax. Staining then occurs more slowly, and only if the embedding medium has not melted or become unduly soft after catting. To avoid this problem, sections are flattened on water no hotter than 45 C and dried overnight at 40 C. Minor technical modifications to the staining procedures are needed. Mercury deposits are removed by iodine, and a 3% solution of sodium thiosnlfate in 60% ethanol is used to remove the iodine from paraffin sections. At room temperature, progressive staining takes 10-20 tunes longer for sections in paraffin than for hydrated sections; at 45 C, this can be shortened to about three times the regular staining time. After staining, the slides are rinsed in water, air dried, dewaxed with xylene, and coverslipped in the usual way. Nuclear staining in the presence of wax was achieved with toluidine blue, O, alum-hematoxylin and Weigert's iron-hematoxylin. Eosin and van Gieson's picric acid-acid fuchsine were effective anionic counterstains. A one-step trichrome mixture containing 3 anionic dyes and phosphomolybdic acid was unsuitable for sections in wax because it Imparted colors that were nninformative and quite different from those obtained with hydrated sections. Advantages of staining in the presence of wax include economy of solvents, reduced risk of overstaining and strong adhesion of sections to slides.     

Suitability of Different Protein A-Gold Markers for Immunogold-Silver Staining in Paraffin Sections

An incubation protocol to immunolabel Lowicryl semithin sections was applied to paraffin probes. To improve the labeling density, colloidal gold complexes of different preparations and sizes were compared. The type of colloidal gold preparation used was found to affect the specificity of the immunostaining. Gold colloid of 5 nm diameter particle size prepared with white phosphorus minimized nonspecific background labeling of β-casein in paraffin embedded sections of the mammary epithelium of pregnant mice. Gold colloids of 5 nm and 9 nm diameter particle size prepared in varying concentrations of tannic acid generated significant nonspecific staining in similar tissue preparations.     

Suitability of Different Protein A-Gold Markers for Immunogold-Silver Staining in Paraffin Sections

An incubation protocol to immunolabel Lowicryl semithin sections was applied to paraffin probes. To improve the labeling density, colloidal gold complexes of different preparations and sizes were compared. The type of colloidal gold preparation used was found to affect the specificity of the immunostaining. Gold colloid of 5 nm diameter particle size prepared with white phosphorus minimized nonspecific background labeling of β-casein in paraffin embedded sections of the mammary epithelium of pregnant mice. Gold colloids of 5 nm and 9 nm diameter particle size prepared in varying concentrations of tannic acid generated significant nonspecific staining in similar tissue preparations.     

不同固定液对豚鼠免疫器官石蜡切片质量的影响

摘要 目的：比较不同固定液对SPF级豚鼠免疫器官的固定效果,为筛选适用于脾脏、淋巴结和骨髓的固定液提供参考。方法：将21只300～350 g雄性SPF级豚鼠随机分为7组,每组3只,摘取脾脏、肠系膜淋巴结、髂骨骨髓,各组分别使用4 %多聚甲醛溶液、10 %中性福尔马林溶液、Bouin''s溶液、Carnoy溶液、Davidson''s溶液、Zenker溶液、Helly溶液固定48 h。髂骨骨髓在固定前浸泡于EDTA脱钙液（pH7.2）中并置于37 ℃孵箱进行脱钙。通过制作石蜡切片并进行HE染色,从切片龟裂程度、细胞形态、染色效果等方面比较7种固定液对切片质量的影响。结果：10 %中性福尔马林溶液固定后的脾脏未出现龟裂,白髓和红髓着色清晰,淋巴细胞形态易辨。Carnoy溶液固定后的淋巴结生发中心明区暗区分明,红蓝染色适中,淋巴细胞形态清晰。4 %多聚甲醛溶液固定后的骨髓着色适中,清晰可辨。结论：常温条件下对组织固定48 h,经HE染色后想要达到理想的切片质量,建议使用10 %中性福尔马林溶液固定脾脏,使用Carnoy溶液固定淋巴结,首选4 %多聚甲醛溶液固定骨髓,如果考虑固定的同时完成脱钙建议选择Bouin''s溶液固定骨髓。     

Application of the Papanicolaou Stain to Paraffin Sections

Routine paraffin sections from tissues fixed either in aqueous formalin, 80% alcohol (with or without 1% trichloracetic acid added), Carnoy's alcohol-chloroform-acetic (6:3:1) and Bouin's fixative were stained as follows: Harris' hematoxylin, 6 min; running water, 2-3 min; ascending grades of alcohol to 95%; orange G, 0.5% and phosphotungstic acid, 0.015% in 95% alcohol, 5 min; 95% alcohol, 2 changes; Papanicolaou's EA36, 2.5 min; dehydration, clearing, and covering in Permount. The results show morphology better than hematoxylin and eosin and the technic is recommended particularly for keratin, which always stains bright orange.     

Postadhesive Technique for Archival Paraffin Sections

We present a postadhesive protocol for adhering paraffin sections of archival material to microscope slides. Appropriately posttreated sections, subsequently processed for immunohistochemistry, remained attached to the slides and were well preserved with no signs of artifacts, such as scratching and shrinkage. The immunohistochemical staining was intense and antigen-specific without nonspecific background. Specific staining intensity was equal to that produced in untreated control sections; however, the latter became partially or fully detached from the slides. The postadhesion protocol may be used with modern techniques and is recommended for reclaiming use of otherwise unsuitable paraffin sections of archival material.     

A Gelatin Fixative for Paraffin Sections

Mayer's albumen fixative, of which the active principle is white of egg, is used almost universally for affixing paraffin ribbons to the slide. About eight years ago the writer's attention was called to a gelatin fixative which has proved to be so superior to albumen that he has used it almost exclusively ever since in the making of a great variety of botanical preparations, and has recommended it to a number of other workers whose experience with it subsequently has been just as satisfactory. The gelatin method was first described by Szombathy¹ and later discussed by Artschwager,² but it does not seem to have received the attention in the literature which its importance deserves. It certainly merits a wide spread use among both botanists and zoologists.     

A Gelatin Fixative for Paraffin Sections

Mayer's albumen fixative, of which the active principle is white of egg, is used almost universally for affixing paraffin ribbons to the slide. About eight years ago the writer's attention was called to a gelatin fixative which has proved to be so superior to albumen that he has used it almost exclusively ever since in the making of a great variety of botanical preparations, and has recommended it to a number of other workers whose experience with it subsequently has been just as satisfactory. The gelatin method was first described by Szombathy¹ and later discussed by Artschwager,² but it does not seem to have received the attention in the literature which its importance deserves. It certainly merits a wide spread use among both botanists and zoologists.     

A Pre-embedding Reaction Method for Localizing NADH Reductase and Succinic Dehydrogenase in Skeletal Muscle

Paraffin wax embedding methods suitable for demonstrating the distribution of enzyme activity in tissues sections are uncommon; most procedures rely on the use of frozen section techniques. This paper describes a system for demonstrating certain enzymes which involves incubation of the tissue with appropriate substrates before a Paramat wax embedding procedure. While it has distinct merits of its own, the procedure is eminently suitable for use where a cryostat is not available; it can also be readily applied to other enzymes and tissues.