冰冻切片技术在植物显微结构和组织化学中的应用

介绍了冰冻切片法研究植物显微结构和组织化学的一般程序。结果表明,冰冻切片过程简单有效且图版清晰,在1d内可获得高质量图片,解决了利用普通石蜡切片观察植物样品需要进行脱水、浸透、包埋操作且耗时长的问题,在植物显微结构和组织化学研究中具有广阔的应用前景。     

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。     

冰冻切片与石蜡切片对乳腺肿瘤的诊断价值研究

目的:分析和比较冰冻切片与石蜡切片对乳腺肿瘤的诊断价值。方法:选取480例新鲜乳腺标本,将其制成冰冻切片以及石蜡切片,根据诊断结果进行对比分析,评价乳腺肿瘤的冰冻切片与石蜡切片的对乳腺肿瘤的诊断价值。结果:经石蜡切片诊断乳腺良性肿瘤277例,占57.71%,良性肿瘤中以乳腺纤维瘤诊断居多;经石蜡切片诊断乳腺恶性肿瘤203例,占42.29%,以乳腺浸润性导管癌居多。冰冻切片诊断乳腺良性肿瘤279例,占58.13%;恶性肿瘤195例,占40.62%;延迟诊断6例,占1.25%。以石蜡切片诊断结果为金标准,冰冻切片诊断乳腺良性肿瘤的准确率为98.56%(273/277),诊断恶性肿瘤的准确率为95.07%(193/203),假阳性率为0.72%(2/277),假阴性率为2.96%(6/203),冰冻切片与石蜡切片诊断乳腺肿瘤的结果具有显著一致性,K值为0.965(P0.05)。结论:冰冻切片与石蜡切片诊断乳腺肿瘤的符合率较高,可作为术中快速病理检测的手段,但该种切片方式存在少量延迟诊断,多与术者操作经验有关,故术中应注重制片过程,提高冰冻切片质量。     

胸腹水脱落细胞学瘤包石蜡切片免疫组织化学染色的应用

胸腹水脱落细胞学涂片做免疫组织化学染色,因具有取材方便、易行,病人痛苦小等优点而适用于临床[1]。脱落细胞学涂片免疫组织化学染色技术最需要注意的问题是背景染色和涂片粘附的牢固程度。因为背景的存在会影响阳性结果的判断,尤其是抗原表达少的弱阳性病例;而涂片涂的不好、厚薄     

免疫组织化学技术检测石蜡切片组织中BCG抗原的体会

为了进一步提高免疫组织化学技术在检测结核杆菌感染的组织中免疫牛型分枝杆菌 (ＢＣＧ)抗原的表达 ,我们收集了2 0例结核病变的组织标本。用不同的抗原修复方法比较和摸索 ,并采用抗酸染色做对照 ,比较抗酸与ＢＣＧ的结果及如何制作出较令人满意的免疫组织化学染色结果得出一点体会 :1 材料和方法1 1　病例 :收集我院手术切除的经病理组织学确诊的结核病变标本 2 0例 ,均经 10 %福尔马林液固定 ,石蜡包埋的组织标本。1 2　使用ＤＡＫＯ ,ＥｎｖｉｓｏｎＳｙｓｔｅｍ二步法试剂盒 ,兔抗牛型分支杆菌 (ＢＣＧ)多克隆抗体。1 3　不同的抗原修…     

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     

防止石蜡切片材料染色时脱落的简易方法

防止石蜡切片材料染色时脱落的简易方法张松林,金芝兰（西北师范大学生物系,兰州７３００１０）ＡＳＩＭＰＬＥＭＥＴＨＯＤＴＯＰＲＥＶＥＮＴＴＨＥＳＰＥＣＩＭＥＮＯＦＰＡＲＡＦＦＩＮＳＥＣＴＩＯＮＴＲＯＭＳＨＥＤＤＩＮＧＤＵＲＩＮＧＳＴＡＩＮＩＮＧ￥Ｚｈａ...     

福尔马林长时间固定石蜡切片免疫组织化学染色体会

许多标本经过福尔马林长时间固定后,其免疫组织化学染色会具有一定的难度,就此,本人进行了如下总结：（1）切片入二甲苯,37℃,两次,各10min,彻底脱蜡。（2）无水乙醇,两次,各10min。（3）95％、90％、80％酒精各5min。     

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     

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.     

Serial Sectioning of Insects with Hard Exoskeleton by Dissolution of the Exocuticle

The method reported here was designed to produce paraffin serial sections as thin as 5 Mm of insects or other arthropods with a hard cuticle. Heads and abdomens of Apis mellifera, Eristalomyia tenax and Tenebrio molitor were fixed with Schaffer's liquid, dehydrated with 80% ethanol, 90% ethanol, two changes of 100% isopropanol (2 hr each) and 12 hr in a 1:1 mixture of paraffin (58 C melting point) at 60 C. They were molded in paraffin after 12 hr of infiltration under a partial vacuum at 60 C. Large body openings of objects were sealed with paraffin to prevent infiltration of solvents.

Thereafter, the outer paraffin was removed manually and with xylene (15 min); the cuticle was rehydrated with 100% isopropanol and 100% ethanol (15 min each). The objects were then treated with Sputofluol (Merck; a mixture of NaOH and NaCIO) until they became white or their colorless endocuticle was stainable with aniline blue WS (C.I. 42755) after rinsing in a 50% acetic acid solution (v/v). They were then dehydrated with 100% ethanol and 100% isopropanol (15 min each) and subsequently re-embedded in paraffin. They were molded, sectioned, stained and mounted as usual.     

Double Embedding Broad Thin Leaves in Paraffin

Paraffin pellets were melted in 24 × 24 × 5 mm stainless steel base molds. Specimens of leaves, 18 × 18 mm, were fixed, dehydrated and infiltrated with paraffin. Two specimens were transferred into molten paraffin on their laminar surfaces in a base mold and moved quickly onto a cold surface to cast them in a shallow block of paraffin. Each block was then scored with a razor blade, broken into two primary blocks, and trimmed to 20 × 9 mm with 5 mm flat edges. Each primary block was immersed upright on its long edge in a 22 × 22 × 20 mm Peel-A-Way® embedding mold containing molten paraffin. The leaf edge was held centrally in the mold while moving the double embedment onto a cold surface. In this secondary block, the leaf specimen stood perpendicular to the sectioning surface in perfect orientation for transverse ribbon sectioning. The two phases of paraffin bonded well.     

Acridine Orange Fluorescence in Paraffin Sections

The technique of staining with acridine orange for fluorescence microscopy of fresh animal and plant cells, chiefly for the detection of ribonucleic acid in the cytoplasm, was brought to a high degree of perfection by Schümmelfeder (1950) and has been developed further by Bertalanffy and Bickis (1956). Its employment for cancer detection in smears was reviewed by Bertalanffy, Masin and Masin in 1956.     

植物材料快速石蜡制片方法

真空干燥箱已越来越广泛地应用于现代生物学研究领域。该文利用真空干燥箱温度和负压的可控制性能,将固定、脱水、透明和石蜡渗透等过程在真空干燥箱中进行,建立起一套可行的植物组织快速石蜡制片方法。结果显示,真空干燥箱的应用加速了多种试剂的渗透速率,提高了切片质量,达到了优化实验步骤、节省实验时间和减少室内有毒化学气体污染的目的。     

Softening Hair with Dithiothreitol (Cleland's Reagent) for Histological Sectioning in Paraffin

Undecalcified embedment of large bone specimens is often challenging. A method is presented here that is suitable for methacrylate embedment of sections of canine vertebrae while retaining the ability to localize tartrate-resistant acid phosphatase and alkaline phosphatase activity. Specimens also retained tetracycline labelling, and sectioned preparations were readily stained with routine bone procedures. A modification of the Bodian silver stain, used for examining the nerves and spinal cord in these specimens, provided a useful stain for canaliculi and cement lines in trabecular and cortical bone. This stain is advantageous when both bone and nerve tissue are of interest, as in spinal fusion studies.     

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.     

Embedding Thin Plant Specimens for Oriented Sectioning

Small plant structures such as small primary roots, filamentous mosses and algae are difficult to orient for sectioning since they become wavy and curl during embedding. A method is described for embedding and orienting tiny plant specimens in a glycol methacrylate resin using self-constructed flat molds. Prior to sectioning, small samples can be oriented in both the longitudinal and the transverse plane. As several samples can be sectioned simultaneously, time-consuming trimming of the blocks is reduced substantially. The efficiency of this technique has been demonstrated using the tiny roots of the model plant Arabidopsis thaliana (L.) Heynh.     

成熟玉米秆组织结构切片方法的比较研究

于玉米成熟期选择健壮茎秆的基部第三节间为材料,采用徒手切片法、冰冻切片法、石蜡切片法、薄切片法等4种方法,比较不同方法的玉米茎秆组织结构切片质量,为研究玉米茎秆结构与其倒伏的关系奠定技术基础。结果表明:徒手切片法是获得成熟玉米秆组织结构切片较为方便、快速的方法,切片面积较大,适合大范围观察统计;冰冻切片法是获得成熟玉米秆组织结构较快的方法,切片面积较小,适合小范围观察;薄切片是获得高质量成熟玉米秆组织结构切片的最好方法,但切片面积太小,适合高倍数显微观察和小范围电镜扫描观察组织结构;石蜡切片不适合作为成熟玉米秆的组织切片方法。研究认为,徒手切片法是最适合成熟玉米秆组织结构观察研究的制片方法。     

高山榕染色体制片优化及核型分析

以高山榕种子的根尖为材料进行染色体常规压片,比较不同预处理方法和解离方法对高山榕染色体制片的影响,以选择最优的压片方法制片并进行核型分析。结果显示,预处理24 h的总体效果优于12 h;3种预处理液的总体作用效果为0.05%秋水仙素>混合液>0.002 mol.L-18-羟基喹啉,综合比较后认为混合液低温4℃处理24 h为最佳预处理方法。解离方法选择1 mol.L-1HCl中60℃水浴2~3 min较为适宜。首次报道了高山榕核型公式为2n=2x=30=22m(2SAT)+6sm+2T,核型不对称系数为61.54%,核型属于Stebbins核型分类中的2C类型。