超声波配合真空技术在石蜡组织块制备中的应用

超声波配合真空技术在石蜡组织块制备中的应用杨金镇我们应用超声波对取材组织进行固定、脱水和透明，然后再真空浸蜡。结果蜡块制备时间明显缩短，切片染色后组织结构和细胞形态清晰。该法是日常外检快速病理诊断的一种新方法。１材料和方法１．１标本和试剂选用宫颈息肉...     

正丁醇替代无水乙醇和二甲苯在制作大鼠全胚切片中的应用 

目的 探索一种以正丁醇替代无水乙醇和二甲苯制作不同胎龄大鼠全胚切片的简便方法。方法 收集胚龄14d（E14）、E16、E18、E20大鼠胚胎，固定及低浓度乙醇脱水后，部分标本按传统方法用无水乙醇脱水和二甲苯透明(无水乙醇-二甲苯法)，另一部分标本用正丁醇脱水、透明(正丁醇法)。所有标本进行石蜡包埋、切片、H     

乙二胺四乙酸联合正丁醇在大鼠胫骨癌痛标本石蜡切片制作中的应用

目的 探索一种简便有效的制作大鼠胫骨癌痛标本石蜡组织切片的方法。方法 取大鼠胫骨癌痛模型组和正常组胫骨标本，使用乙二胺四乙酸（EDTA）加温脱钙联合正丁醇取代传统强酸脱钙、纯酒精脱水和二甲苯透明，制作石蜡切片后HE染色观察。结果 两组大鼠胫骨癌痛标本形态结构均保存完整。与传统方法相比，制片步骤简单，脱水和透明时间较为灵活，蜡块切割性能较好。结论 EDTA与正丁醇的联合应用，是大鼠胫骨癌痛标本石蜡组织切片制作适宜和可靠的方法。     

微波配合真空技术在石蜡组织块制备中的应用

微波配合真空技术在石蜡组织块制备中的应用杨金镇我们应用微波对取材组织进行固定、脱水和透明，然后再真空浸蜡。结果，组织块从固定到浸蜡在３０～７５分钟完成，且单例出片时间缩短到７０～１５０分钟，切片染色后组织结构和细胞形态清晰艳丽同常规方法，是病理科日常...     

人胚胎肠标本石蜡制片方法的探讨

石蜡切片因操作简单,易保存等优点而成为现代组织学、胚胎学、病理学等医学科学研究领域最常使用的一种方法。石蜡切片制作包括取材、固定、脱水、透明、浸蜡等许多环节,每一环节都直接影响石蜡切片制作结果。目前动物标本及     

石蜡包埋对兔肝、肾组织块的收缩影响

本实验是调查石蜡包埋对兔肝、肾组织块的收缩影响。用米尺和下端封闭、带刻度的玻璃管对新鲜组织块到石蜡包埋全过程中组织块长、宽、厚及体积的变化进行测量并进行统计学处理。结果表明从固定、脱水、透明到浸蜡每一步骤都使组织块发生了收缩 ,而浸蜡使其收缩最为明显 ,肝组织块收缩了 5 0 .0 0 % ,肾组织块收缩了 5 8.13%。因此包埋过程要尽可能的缩短时间 ,尤其浸蜡要最大限度地降低温度和缩短时间。石蜡切片在镜下不能精确地反应正常组织结构的大小。     

环保型BT生物组织透明剂替代二甲苯在病理组织透明中的应用及比较

临床送检的病理组织标本经病理医师取材后,需要进行固定、脱水、透明、浸蜡等处理才能制作石蜡切片,透明效果的好坏直接影响到石蜡制片的质量.二甲苯是病理组织透明最常用的试剂,但二甲苯具有毒性,不利于病理工作人员的身体健康和环境保护.我们使用环保型BT生物组织透明剂替代有毒的二甲苯应用于病理组织透明,制作病理切片,效果满意,现介绍如下.     

硬质酸石蜡包埋组织观察

本实验采用取10％甲醛、Bouin液分别固定新鲜兔的肾上腺和肺，常规脱水。取硬质酸按比例加入石蜡。置入恒温箱内溶解，然后放入彻底脱水组织，取消用二甲苯和石蜡完成对组织透明、浸蜡。用石蜡包埋、常规切片、染色。显微镜下观察；用甲醛固定的组织切片符合教学用片要求，而用Bouin液固定的则存在大量针尖大小的颗粒。     

影响淋巴结HE制片因素的探讨

淋巴瘤因其组织结构致密、细胞丰富等特点,使组织在脱水、透明、浸蜡、切片、染色等方面都受到一定的影响,导致病理诊断困难。作为一名合格的病理医师,要求懂得组织的石蜡包埋和HE染色的机制,熟练掌握石蜡切片和HE染色技术的操作并能够制出一张优质的病理玻片标本。本实验从石蜡切片和HE染色的机制出发,重点探讨淋巴结HE制片的注意事项。     

自动组织脱水机应用体会

随着病理技术的发展,组织的人工脱水已逐步被组织脱水机代替,其最大特点是能利用非上班时间进行处理组织,并也能保证组织脱水、透明和浸蜡效果。由于病理标本种类较复杂,组织大小不一等因素,故要使组织处理达到最佳效果,也是不易掌握的。我科近年在使用自动组织脱水机过程中,对脱水剂、透明剂、浸蜡剂更换时间和组织数量处理数量关系进行了改进,从而提高石蜡切片质量。１　试剂１－１　固定液的选择　为了给组织切片和染色提供有利条件,需根据待固定组织的组织结构或细胞成分,选择不同的固定剂。我们根据固定剂的性质和条件而选择…     

A Comparison of Lectin Binding to Frozen,Formalin Fixed Paraffin Embedded,and Acid Decalcified Paraffin Embedded Tissues

Abstract

A comparison was made between lectin binding to frozen, formalin fixed paraffin wax embedded, and to formic acid treated, paraffin wax embedded tissues, using three lectins UEA I, PNA, SBA. The binding to various rat tissues was assessed with a multiple layer immunoperoxidase staining method. Except for an increase in binding to some tissues, there was little difference between the acid treated and non-acid treated formalin fixed, paraffin embedded tissues. With several tissues, differences between frozen and paraffin embedded specimens suggested that care should be exercised in the choice of tissue processing if lectin histochemistry is to be undertaken. (The J Histotechnol 11:223, 1988.)     

人内耳骨迷路的组织发生

目的 :了解人胚胎内耳骨迷路的组织发生。方法 :取因故人工流产的胚胎 42例 ,将内耳作石蜡包埋 ,连续切片 ,光镜观察。扫描电镜标本用冷冻割断法 ,观察其割断面的表面结构。结果 :胚胎第 6周蜗管周围的间充质聚集 ,7- 8周变为初软骨 ,9- 12周分化为真性软骨 (透明软骨 ) ,14周骨化开始 ,2 5周骨化完成。结论 :骨迷路的形成是以软骨内成骨的方式进行的     

人胚胎期垂体的形态学研究

将各胚胎期垂体分别测量后速入Bouin或Helly液固定,石蜡包埋,将连续切片进行特殊染色,在光镜下观察并记述了腺垂体和神经垂体的早期发生过程及内部构筑。我们将腺垂体的发生过程分为原基变形期、增生期和增生分化期。实验中记录了各种阳性细胞出现时间及百分值,并且將各种数据进行了统计学处理。文末对垂体发生和分化的动力学问题进行了讨论。     

Improved double immunohistochemical staining method for cryostat and paraffin wax sections, combining alkaline phosphatase anti-alkaline phosphatase and indirect immunofluorescence.

AIMS--To develop and immunohistochemical staining method for cryostat and paraffin wax sections so that two different antigens in the same section of tissues could be detected by combining immunoenzyme and immunofluorescence techniques. METHODS--This double immunohistochemical staining method combines alkaline phosphatase-anti-alkaline phosphatase (APAAP) using New Fuchsin as a chromogen and indirect immunofluorescence. RESULTS--APAAP staining for one antigen of this double immunohistochemical staining method was observed under bright field conditions alternating with immunofluorescence for another antigen under ultraviolet light. The double exposed photograph of both easily identified the two signals within the same cell. CONCLUSIONS--This double immunohistochemical staining method can overcome the disadvantages of any masking effect of the double immunoenzymatic methods and the background problems of double immunofluorescence method especially when applied to paraffin wax sections. It also permits good morphological identification of the doubly stained cells which may be of crucial importance in studies on pathology specimens.     

结扎大鼠胸导管诱发胰组织结构的形态学变化

目的 观察胰淋巴淤滞动物模型胰组织结构变化和胰淀肽沉积情况,探讨淋巴淤滞对胰组织细微和超微结构的影响. 方法 10月龄大鼠20只,采用腹部结扎大鼠胸导管,建立胰淋巴淤滞动物模型,造模后6个月取材,部分胰组织经石蜡包埋切片,行HE和刚果红染色;部分经冷冻切片,行免疫组织化学染色和光镜观察;部分标本进行透射电镜样品制备和观察. 结果 HE和刚果红染色切片光镜观察显示,胰腺小叶间隙增宽,呈明显的结缔组织增生、脂肪堆积;胰岛淡染或着朱红色,组织间隙明显增宽.免疫组织化学染色切片光镜观察显示,在胰岛及其周围,呈现胰淀肽强阳性棕褐色着色反应.超薄切片透射电镜观察显示,胰腺小叶间隙增宽,可见血管和扩大的淋巴管;胰岛细胞间隙增宽,细胞间隙内可见大量脂滴样物质和胶原原纤维样结构.结论 胸导管结扎可导致胰淋巴引流障碍,引起胰淋巴管扩张,结缔组织间隙增宽,脂肪堆积,胰岛细胞间隙扩大,胰淀肽沉积等,这些结构变化可能影响胰岛的功能.     

Feasibility of in situ hybridisation with chromosome specific DNA probes on paraffin wax embedded tissue.

The feasibility was studied of in situ hybridisation using chromosome specific DNA probes on paraffin wax embedded normal and malignant tissues from different organs. Both isolated nuclei and 5 microns sections were used in in situ hybridisation experiments with biotinylated repetitive DNA probes specific for the centromeric regions of chromosomes 1 and 17. The hybridisation results were visualised with peroxidase-diaminobenzidine. The optimal pretreatments with sodium thiocyanate and pepsin were experimentally defined for the different tissues. Although interphase cytogenetics on paraffin wax embedded tissue is possible, the results indicate that it has its limitations, compared with investigations on fresh tumour tissue.     

New insights into the morphogenesis of the gubernaculum testis and the inguinal canal

There is no consensus about the role of the gubernaculum testis (GT). Nineteen human embryos (Carnegie stages 15–23), 36 fetuses (9 weeks to term), and eight neonates were examined. All the embryos and 25 fetuses (from weeks 9–16) were processed for paraffin wax histology and serially sectioned at 10 µm thickness. The remaining 11 fetuses and the eight neonates were fresh specimens that were dissected under a surgical microscope. The GT appeared during the embryonic period (stages 17–23) with a horseshoe‐shaped mesenchyme condensation of the superior concavity, which was observed in four different topographical regions sequentially through development. The GT was not attached at either of its ends in any of the specimens, not to the gonad or in the scrotal sac. The inguinal canal differentiates around the inguinal portion of the GT during the late embryonic period. After stage 21, the GT appears enveloped by an evagination of the peritoneal cavity. It has few striate muscular fibers and vessels. Although the GT does not appear to have the role traditionally attributed to it, it is still an essential structure and indirectly facilitates the descent of the testes. It contributes to the formation of the inguinal canal and then forges the pathway that the testes will subsequently take through the inguinal canal as they migrate from the abdominal cavity into the scrotal sac. Clin. Anat. 30:599–607, 2017. © 2017 Wiley Periodicals, Inc.     

Demonstration of multiple HPV types in normal cervix and in cervical squamous cell carcinoma using the polymerase chain reaction on paraffin wax embedded material.

The prevalence of human papilloma virus (HPV) types 6, 11, 16 and 18 was investigated using the polymerase chain reaction on formalin fixed, paraffin wax embedded material in 19 cases of cervical squamous cell carcinoma and in 10 normal cervices. HPV DNA was detected in 16 of 19 carcinomas, with multiple types present in 11 of these. HPV 16 or 18, or both, were present in all cases in which HPV was shown. Six of 10 cases of normal cervix contained HPV; five of these contained two or more HPV types, including HPV 16 or 18, or both. This study shows the feasibility of using the PCR on paraffin wax embedded material and indicates a high rate of carriage of multiple HPV types in both normal and neoplastic cervix.     

Immunohistochemical demonstration of c-erbB-2 oncoprotein in gastric adenocarcinoma: comparison of cryostat and paraffin wax sections and effect of fixation.

AIMS--To investigate the effects of fixation on the immunohistochemical demonstration of c-erbB-2 oncoprotein using paraffin wax and cryostat sections; to compare c-erbB-2 expression in non-neoplastic and neoplastic gastric tissues. METHODS--Adjacent blocks of tumour and non-neoplastic tissue from four gastrectomy specimens were put into a panel of 10 fixatives including acetone, B5, Bouin's fluid, Carnoy's fluid, buffered formalin, formol dichromate, zinc formalin, 4% paraformaldehyde, periodate-lysine-paraformaldehyde (PLP) and periodate-lysine-paraformaldehyde-dichromate (PLPD) before embedding in paraffin wax for sectioning. Similar tissue blocks were snap frozen and cryostat sections were postfixed in these fixatives, either alone or in combination, before immunostaining. RESULTS--In paraffin wax embedded sections the best fixative was PLP, and in frozen tissues the best results were obtained after fixation of cryostat sections in buffered formalin followed by cold methanol and acetone. Applying these fixatives to samples from a further 16 gastrectomy specimens, strong membrane staining of c-erbB-2 protein was found in the tumour in eight of 16 cases (50%) using paraffin wax sections, and staining was stronger in the better differentiated carcinomas. For frozen tissues, positive membrane staining was found in all gastric adenocarcinomas, but differential staining intensity associated with tumour differentiation could not be detected. CONCLUSIONS--These results indicate that fixation and paraffin wax embedding affect the results of immunohistochemical demonstration of c-erbB-2 in gastric cancer. The choice of fixative is critical in the demonstration and evaluation of c-erbB-2 protein expression by immunohistochemistry in gastric carcinomas. Staining results also vary depending on whether frozen or paraffin wax embedded tissues are studied.