塑料不脱钙、石蜡包埋、明胶浸泡法对骨组织切片的比较

骨组织内含有大量的钙盐和无机盐,如不脱钙切片,常给骨组织切片和染色造成非常大的困难,直接影响实验结果。作者根据研究的需要,对塑料包埋不脱钙、石蜡包埋、明胶浸泡３种方法切片及染色进行比较,以便找出一种最佳切片方法。１　材料与方法１－１　塑料包埋组　所用材料是我所研究生课题实验材料。新西兰大耳白兔１０只,体重２－２～２－５ｋｇ,♀、♂不限。取双侧尺桡骨１－５ｃｍ保留周围少量肌肉。１０％中性福尔马林固定。梯度乙醇脱水。甲液：甲基丙稀酸甲酯（ＭＭＡ）７５ｍｌ,邻苯二甲酸二丁酯（ＤＢＰ）２５ｍｌ。乙液：Ｍ…     

塑料包埋不脱钙骨组织玻璃刀切片技术

一、材料和方法1．取材固定：用307－6型台式牙钻车，换上金钢砂片切取长约1cm；宽约0．5cm；厚约0．3cm骨组织，用7O％乙醇固定3～5天。2．脱水透明：用80％、90％、100％乙醇脱水、二甲苯透明，每级各二次，每次24小时。3．浸透液：I液：甲基丙稀酸甲酯（MMA）75ml，邻苯二甲酸二丁酯（DBP）25ml。Ⅱ液：MMA75ml，DBP25ml。过氧化苯甲酸1g。Ⅲ液：MMA75ml，DBP25ml，过氧化苯甲酸2．5g。以上三液，配时均需搅拌数小时，在搅拌机上充分搅匀。Ⅰ、Ⅱ、Ⅲ液各浸透36小时。Ⅲ液也为包理液。4．包埋：将青霉素小瓶中滴入2ml包埋液，40…     

电镜树脂包埋液配方的选择与应用

用于透射电镜样品的包埋液 ,数年来一直延用几种固定的配方 ,即常用的81 2、国产树脂 6 1 8、Spurr等等 ,各种包埋剂各有其优缺点。Spurr包埋剂 :粘度低 ,浸透容易 ,包埋聚合时间短 ,硬度及韧性均很好 ,经免疫标记的样品 ,常用此液进行包埋。另外还可用于难以浸透、硬度高、致密性较高组织。但此包埋液价格昂贵 ,不宜作为常规的包埋液来使用。81 2包埋液 :粘度低 ,韧性好 ,易切出较大的超薄切片 ,但组织包埋块易吸潮 ,不易保存 ,包埋液的价格也较高。国产树脂 6 1 8:价格便宜 ,购买方便 ,组织块不易吸潮 ,保存方便 ,切割性能较好。但此包埋…     

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。     

从OCT包埋冰冻组织提取组织RNA和检测病毒RNA

目的探讨如何从OCT包埋冰冻组织提取组织RNA和检测病毒RNA。为临床有限标本的保存和利用提供一简便的方法。方法用汉坦病毒感染的小鼠组织标本，经质量浓度25％蔗糖磷酸缓冲液固定OCT包埋，在-20℃冰箱中保存2年后，对冰冻的OCT包埋的感染组织标本进行RNA提取，用凝胶电泳、紫外分光光度计及RT—PCR等方法检测组织总RNA和病毒RNA。结果组织经质量浓度25％蔗糖磷酸缓冲液固定OCT包埋冰冻保存后，从中提取的RNA仍保持较好的完整性，可扩增出较长片段的病毒RNA。结论OCT包埋的冰冻组织标本不影响提取组织RNA并检测病毒RNA。     

蓝光损伤小鼠视网膜感光细胞功能及病理改变

目的:改为研究蓝光损伤模型中小鼠视网膜感光细胞的功能及病理改变。方法:采用6~8周龄BALB/c小鼠28只,雌雄不限,随机分为实验组及对照组各14只,实验组连续蓝光照射14 d,用罗兰电生理仪观测全部小鼠视网膜电图波幅后处死小鼠,小鼠视网膜感光细胞层的组织病理学改变经固定、包埋、苏木素-伊红(HE)染色后进行光学显微镜观察,超微结构改变经固定、包埋、铅铀双染后进行透射电子显微镜观察,感光细胞层细胞凋亡改变经固定、包埋、TUNEL试剂盒染色后光学显微镜观察。结果:实验组小鼠视网膜电图Rod波形中b波及Max波形中a、b波波幅均较对照组明显下降(P0.05);HE染色后,实验组小鼠视网膜感光细胞层厚度值较对照组明显降低(P0.05),感光细胞核数量减少、排列紊乱;透射电子显微镜下,实验组小鼠感光细胞核内染色质分布不均匀,可见核固缩、核碎裂,内节内线粒体肿胀,可见空泡,外节膜盘部分叠状结构解离、消失;TUNEL染色后,实验组小鼠感光细胞层内见多量棕黄色免疫反应阳性产物沉积。结论:小鼠经蓝光照射后可发生视网膜功能下降,提示这与感光细胞的病理学和超微结构改变以及感光细胞凋亡有关。     

甲醛固定石蜡包埋食管鳞状细胞癌组织蛋白提取方法的比较

目的 探讨从甲醛固定石蜡包埋(FFPE)食管鳞状细胞癌(ESCC)组织中高效率提取蛋白质的方法.方法 6种裂解液和100℃、105℃两种条件分别提取8例甲醛固定石蜡包埋食管鳞状细胞癌组织蛋白,Bradford法测定蛋白浓度、十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)、Western blotting和免疫组...     

c—erbB—2蛋白和HBsAg在肝细胞癌与癌旁组织中的表达及相互关系

ｃ－ｅｒｂＢ－２蛋白和ＨＢｓＡｇ在肝细胞癌与癌旁组织中的表达及相互关系程瑞雪冯德云沈明颜亚辉一、材料与方法取我校附属湘雅医院外科手术肝癌（ＨＣＣ）标本４０例，均附有癌旁组织；尸检正常肝组织１０例。所有标本经１０％福马林固定，石蜡包埋，５μｍ厚连续切片...     

肾穿刺组织固定液的改进

肾穿刺组织固定液的改进郑智勇，李京榕肾穿刺组织固定液通常采用缓冲福马林、ＦＡＡ液（福马林－酒精－冰醋酸）、Ｄｕｂｏｓｃｑ－Ｂｒａｓｉｌ液（福马林－酒精－苦味酸－冰醋酸）或福马林升汞液。第一种固定液使肾小管细胞浆收缩及刷状缘磨损；中间两种固定液对肾小管...     

改进石蜡包埋法作大鼠全肺2μm连片一醛品红与Giemsa染法比较探讨

大白鼠肺组织切片易碎,不易制成大片完整的2μm连片。作者经反复实践,在包埋、切片、染色等方面作了一系列的改进,在制成大白鼠全肺2μm连片,用醛品红、光绿对比染色与Giemsa染色比较,并获得较满意的结果。兹将方法介绍如下。 1.将大白鼠全肺固定于10％中性缓冲甲醛液内2天。更换新鲜固定液1次。     

Fetal calf serum versus human serum: ultrastructural evaluation of protein support influence on human ovarian tissue cryopreservation

The aim of this study was to examine the effect of the different protein supports in the cryopreservation solution on improving human ovarian tissue preservation after frozen-thawed procedures. Biopsies of ovarian cortical tissue were obtained from 14 subjects. All specimens were cryopreserved using a slow freezing/rapid thawing method in a solution consisting of propanediol and sucrose in different proportions of 3 protein supports: 30% human serum (HS) (solution A), 20% HS (solution B), or 20% fetal calf serum (solution C). After thawing, 191 follicles and a total of 70 samples were analyzed using transmission electron microscopy (TEM). The post-thaw preservation rate of the follicles in solution A was significantly higher with respect to solution C (p < 0.05). Unlike the follicles, the stromal cell morphology was not affected by any of the solutions investigated. By comparing stromal morphology and the patient age, it was found that HS better preserved the tissue in patients over 20 years of age with respect to younger ones, which showed a wider variability in ovarian preservation. TEM evaluation showed that 30% HS is more suitable for human ovarian tissue cryopreservation, and research should be focused on defining cryopreservation protocols specific to young patients.     

A method for the differential staining of muscle and connective tissues from organs and their complexes]

The method of differential staining of muscles and connective tissue in the genitourinary organs (urinary bladder, ureters, renal pelvis) and gastrointestinal tract is suggested. The method consists of the intermittent use of known tissue van Gieson dyes (picric acid and fuchsin). When applied at pH = 7.2-8.0 (phosphate or citrate buffer) they stain the connective tissue in red and muscles in gold-yellow colour. A saturated solution of picric acid was mixed at equal proportions with water-free glycerol. Fuchsin was taken as 0.3% buffered solution or as 1% solution applied on the organ intermittently with picric acid. The organs, before staining, are fixed in 0.3 alcoholic solution of carbonic acid.     

Methods of treating tissues with subsequent embedding in paraffin

Two methods of tissue processing with subsequent embedding into paraffin are proposed. In the first method a mixture of 70% of gum turpentine and 30% oleoresin is used as the main solution. In the second one a mixture of gum turpentine, castor oil and cedar balm is used as the main solution. The treatment of the tissue pieces takes 23 hours. These methods permit avoiding the artifacts connected with tissue dehydration and obtaining histological slides of a high quality.     

Buffered formalin is the superior fixative for the detection of HPV DNA by in situ hybridization analysis.

In situ hybridization is used commonly for detection of human papillomavirus (HPV) DNA. There is little information, however, on whether the detection of HPV DNA by in situ hybridization can be affected by the way in which the tissue is fixed. To address this question, the authors compared the hybridization signal using this technique under low stringency conditions for several genital condylomata containing HPV 6 or 11 that were randomly subdivided and fixed in various fixatives for 16 hours. In all cases, the largest proportion of cells with koilocytotic atypia that had detectable HPV DNA was in buffered formalin-fixed tissue (80%), followed by tissue fixed in unbuffered formalin (70%), Hartman's solution (40%), and Bouin's solution (10%). After a high stringency wash, the greatest decrease in the overall hybridization signal was with tissue fixed in Bouin's solution; a minimal decrease was noted with tissue fixed in buffered formalin. Fixation in Bouin's solution for 2 hours gave in situ hybridization results comparable with buffered formalin fixation but with poorer cytologic detail. It is concluded that, of the fixatives studied, buffered formalin is superior for the detection of HPV DNA by in situ hybridization analysis.     

豚鼠肠系膜淋巴管平滑肌细胞的培养

目的 :培养豚鼠肠系膜淋巴管平滑肌细胞。方法 :肠鼠肠系膜淋巴管用胰蛋白酶消化二次 ,第一次消化 15min ,除去肠系膜、淋巴管外膜等组织 ,第二次消化中膜的平滑肌 ,3 0～ 45min后 ,吸出未消化的组织 ,获得平滑肌细胞。常规培养、传代。结果 :培养细胞经光镜、电镜和荧光显微镜可观察到平滑肌的典型的形态和结构。结论 :所培养细胞为平滑肌细胞 ,此法可应用于管径较小的淋巴管和血管平滑肌细胞的培养。     

A method to fix lipids for staining fat embolism in paraffin sections

AIMS: To develop a method to preserve lipids in formalin-fixed tissues for staining in paraffin sections, and to illustrate its use in lung and brain of a fat embolism case, and in examples of fatty liver and atheroma. METHODS AND RESULTS: A saturated solution of linoleic acid in 70% ethylene glycol was prepared and tissues were exposed to this for 3 days at 56 degrees C. These tissues were treated with 2% chromic acid at 4 degrees C for 24 h followed by 24 h in 5% sodium bicarbonate, with appropriate rinsing between solutions. Paraffin sections of these tissues were stained with a lipid-soluble dye such as Oil Red O. Examples of fat embolism, fatty liver, and atheroma were shown photographically as illustrations of expected results. CONCLUSIONS: The demonstration of fat embolism with good quality tissue detail is made practical by the method, which is convenient and inexpensive. The method appears to be generally applicable to tissue lipids of various sorts, as exemplified by adipose tissue, fatty liver, and atheroma.     

Marking excision margins of surgical specimens by silver impregnation

Marking excision margins of surgical specimens by silver impregnation has several advantages over commonly used Indian ink: during the slicing the tissue preserves its natural color, the staining is permanent, and the pigment does not smudge over cutting surfaces. The pigment is clearly visible in tissue sections. The tissue specimen is shortly dipped into a 10% water solution of argent nitrate (AgNO3 with HNO3). After slicing, the tissue specimens are developed in common black & white developer for several seconds and paraffin processed as usual. The method is suitable for formaldehyde fixed as well as fresh tissue specimens.     

A rapid and simple method of purification of Toxoplasma gondii trophozoites originating from tissue culture for use in the indirect immunofluorescent antibody test

A simple and convenient purification method for Toxoplasma trophozoites from tissue culture cells by density centrifugation is described. Separation of cells treated with formaldehyde is achieved by means of a Percoll solution with a density of 1.056 g/ml permitting removal of more than 99% of the cells and cell debris from the tissue culture. Thus highly purified trophozoites of Toxoplasma gondii raised in tissue cultures become available for serological tests, particularly for the indirect immunofluorescent antibody test.