振荡法在组织石蜡制片中的应用

本文成功地把振荡法应用于组织石蜡制片中,采取鸡的各种组织,用常规、加温、振荡三种方法进行固定、脱水、透明和浸蜡,然后常规切片染色试验比较。结果表明,用振荡法从固定到浸蜡仪用2小时20分钟,提高功效10倍以上。具有传统方法所不及的简便、快速、切片薄及质量好等优点,尤其适用于临床病理检验的快速诊断。完全可以代替一般的快速石蜡切片和冰冻切片。     

18℃恒温下,石蜡切片制作过程的脱水透明最佳时间探索

目的:18℃恒温下,探索石蜡切片制作过程的脱水透明最佳时间,为组织切片技术提供可靠的理论依据,以使作出高质量的石蜡切片.方法:将固定好的肝、脾、心、肾组织按组织类型随机各分为六个实验组(n＝6),每组5块组织,制定六组脱水时间段(min):50、60、75、90、105、120,脱水采用的乙醇.每类组织自实验一至六组在浓度70％、80％、95％I、95％ II各级乙醇中的脱水时间,分别为所制定的六组脱水时间段,另外每组在100％Ⅰ和100％ Ⅱ的乙醇中脱水时间各为30 min,透明时间在二甲苯中肝、脾均为65 min;心、肾均为85 min.观察每类组织的各实验组在不同的时间段进行的脱水透明实验、将所得出的透明结果进行比对.实验制作重复五次,实验程序在18℃电热恒温箱中进行.结果:18℃恒温下,组织的最佳脱水时间在上行梯度各级浓度的乙醇中,肝、脾75 min;心、肾90 min,透明时间在二甲苯中肝、脾65 min;心、肾85 min.结论:在本实验的探索实践中体会到,好的脱水透明效果取决于组织块的大小、实验温度、脱水透明时间等,该研究得出的结果:”18℃恒温下,石蜡切片制作过程的脱水透明最佳时间”,有其有效性、可重复性、可行性,在石蜡切片技术领域可以推行应用.     

正庚烷代替二甲苯在拟南芥花药石蜡切片中的应用

二甲苯是应用最广的一种透明剂,常用于石蜡切片中的脱蜡和透明,但由于其对人体危害大,广大科研工作者一直在探寻更加安全的二甲苯代替物。以不同时期的拟南芥花药为试材,脱蜡和透明均由正庚烷代替二甲苯完成,其余步骤与常规石蜡切片相同。结果显示,正庚烷透明力强而迅速,既能与乙醇、封藏树胶混合,又能溶解石蜡,可作为合适的透明剂和脱蜡剂;组织经正庚烷处理后物理性质良好,浸蜡充分,切片时软硬适中,容易连续切片,而且厚薄均匀,摊片时能在42℃水中自然展开,平整而无皱褶;经甲苯胺蓝染色,细胞着色好,核质分明,结构清晰,组织形态保持完好,与二甲苯作透明剂的切片效果无明显区别。因此,正庚烷可以作为一种新的、相对安全有效的二甲苯替代物,应用于植物组织切片中。     

微波快速石蜡切片技术及其在肾穿刺活检中的应用

肾穿刺活检技术推动了肾脏病理的迅速发展,使肾脏病理学整体知识不断更新和提高,为临床诊断和治疗提供了大量的依据.由于肾脏活检组织标本小、切片薄,在固定、脱水、透明、浸蜡、切片及染色等方面均与常规石蜡切片有所不同,其技术要求较高.肾穿刺石蜡切片时间较长,相对制约了急性肾功能衰竭、急进性肾炎、肾移植急性排斥反应等急重症的快速诊断.     

基于改良石蜡切片技术的飞蝗胚胎浆膜表皮发育模式及形态变化观察

【目的】通过优化飞蝗Locusta migratoria胚胎石蜡切片制备技术,研究飞蝗胚胎发育时期浆膜表皮变化规律。【方法】通过优化飞蝗胚胎固定前处理、洗涤、脱水和透明等步骤,改良胚胎期石蜡切片制备技术;制备飞蝗胚胎发育第3-10天的石蜡切片,通过HE染色观察浆膜表皮发育规律;通过制备飞蝗发育第6-8天从卵孔端至卵尾端的纵切切片,观察飞蝗胚胎转旋时期的形态变化。【结果】飞蝗胚胎期石蜡切片技术优化后的操作步骤为:固定前Na Cl O预处理和打孔、洗涤30min、脱水30 min和透明30 min。通过此方法可获得表皮结构完整且清晰的胚胎期组织切片。在30℃条件下,飞蝗胚胎浆膜表皮及几丁质沉积形成于胚胎发育的第5天,在第8天时开始降解。飞蝗胚胎的转旋发生在第6-7天,伴随着浆膜和浆膜表皮的形态变化。【结论】本研究优化了飞蝗胚胎石蜡切片制备步骤,揭示了飞蝗胚胎浆膜表皮及胚胎转旋的发育模式,为昆虫胚胎发育的研究提供了基础资料。     

生物组织石蜡切片的不脱蜡染色程序

石蜡切片法是生物学中应用最广泛的一种研究技术,沿用至今已一百余年,形成了一个基本固定的操作程序,为人们所普遍采用。常规的石蜡切片的染色,必须首先用二甲苯将石蜡溶去(即脱蜡),然后经下行的梯度酒精将切片下降至水(水化),用染料的水溶液进行染色;染色后又经上行梯度酒精逐级脱水,经二甲苯透明后用树胶封藏。此染色程序不仅手续较烦琐,而且切片在染色后脱水时容易褪色以及在此过程中组织里一些孤立分散的细胞如孢子、     

脂肪组织石蜡切片制作方法探讨

目的探讨脂肪组织石蜡切片制作方法,改善和提高其切片质量。方法将送检的脂肪组织切开固定于中性甲醛液中;取材后将标本放入脱水机中,调整脱水机设置程序,在常规方法的基础上适当增加组织再固定及脱水、透明时间;包埋时采取挤压组织等改进方法。结果通过改进和调整后制作的脂肪组织蜡块平整无塌陷,石蜡切片优良,厚薄均匀,完整,无空洞、无挤压、镜下组织结构完整清晰,细胞无挤压、重叠等现象。细胞形态清晰,着色均匀艳丽。结论通过上述方法的改进,能尽可能多地减少脂肪组织的脂滴,增加脂肪组织的硬度,使组织和石蜡能很好的融合在一起,有利于切出完整的厚薄均匀的石蜡切片,提高制片质量。     

用石蜡切片法制作内耳切片

用含盐酸的Ｂｏｕｉｎ氏液固定、脱钙软化，延长高浓度酒精的脱水时间，恰当掌握透明时间，注意耳蜗及半规管的包埋方向等方法制作内耳石蜡切片，获得了该器官主要组织结构较理想的切片。     

人体及动物组织H.E染色石蜡切片法的技术改进

报道人体及动物组织石蜡切片方法的全面革新技术。以组织块的固定、苏木精染色和脱水同时进行为主要技术特征,改进了某些操作细节,获得了良好的效果。     

石蜡切片法中染色技术的改良

石蜡切片是观察动植物组织最直接有效的方法之一。本实验对水稻品种9311的叶片进行切片并用1%的番红—乙醇染液和1%的苯胺蓝—乙醇染液进行染色,结果显示此改良方法得到的组织比传统石蜡切片更完整,染色的效果比传统石蜡切片更清晰,并且利用此方法对水稻叶片进行制片,可以大量缩短制片后期复水及脱水的时间,提高工作效率。     

Chloroplast DNA extraction from herbaceous and woody plants for direct restriction fragment length polymorphism analysis

The technique described here is a fast and simple method of extracting chloroplast DNA (cpDNA). It overcomes the need for differential centrifugation using density gradients. The leaves do not have to be kept in the dark and lyophilized before extraction, but lyophilization is still possible. The chloroplasts are specifically lysed in a cell extract of leaves, using a non-ionic detergent. After isolation by centrifugation, the cpDNA is purified by the combined action of proteolytic enzymes and detergents, followed by the elimination of proteins using a mixture of chloroform and isoamyl alcohol. This method provided good quality restriction profiles for all species analyzed.     

A Silver Impregnation Technique for Normal Axons in the Human Central Nervous System for Celloidin and Epon Sections, with Substitutes for Soft Tap Water

An improved silver technique has been developed for human CNS axons in sections from celloidin blocks that resist impregnation because of prolonged storage in alcohol. This method also gives consistently good impregnation of recently fixed material, and thus is suitable for routine use. Slightly modified, the method is also successful with osmicated Epon embedded sections. The quality of silver impregnation in methods using tap water in the reducing solutions varies in different laboratories. Having established that hard water is essential, substitutes for soft water were sought and found.     

A silver impregnation technique for normal axons in the human central nervous system for celloidin and epon sections, with substitutes for soft tap water

An improved silver technique has been developed for human CNS axons in sections from celloidin blocks that resist impregnation because of prolonged storage in alcohol. This method also gives consistently good impregnation of recently fixed material, and thus is suitable for routine use. Slightly modified, the method is also successful with osmicated Epon embedded sections. The quality of silver impregnation in methods using tap water in the reducing solutions varies in different laboratories. Having established that hard water is essential, substitutes for soft water were sought and found.     

Born Normalization for Fluorescence Optical Projection Tomography for Whole Heart Imaging

Optical projection tomography is a three-dimensional imaging technique that has been recently introduced as an imaging tool primarily in developmental biology and gene expression studies. The technique renders biological sample optically transparent by first dehydrating them and then placing in a mixture of benzyl alcohol and benzyl benzoate in a 2:1 ratio (BABB or Murray s Clear solution). The technique renders biological samples optically transparent by first dehydrating them in graded ethanol solutions then placing them in a mixture of benzyl alcohol and benzyl benzoate in a 2:1 ratio (BABB or Murray s Clear solution) to clear. After the clearing process the scattering contribution in the sample can be greatly reduced and made almost negligible while the absorption contribution cannot be eliminated completely. When trying to reconstruct the fluorescence distribution within the sample under investigation, this contribution affects the reconstructions and leads, inevitably, to image artifacts and quantification errors.. While absorption could be reduced further with a permanence of weeks or months in the clearing media, this will lead to progressive loss of fluorescence and to an unrealistically long sample processing time. This is true when reconstructing both exogenous contrast agents (molecular contrast agents) as well as endogenous contrast (e.g. reconstructions of genetically expressed fluorescent proteins).Open in a separate windowClick here to view.^{(87M, flv)}     

Hydrogels for an accommodating intraocular lens. An explorative study

In this study it was investigated whether hydrogels could be used for an accommodating lens. The requirements of such a hydrogels are a low modulus, high refractive index, transparency, and strength. Since conventional hydrogels do not possess this combination of properties, a novel preparation method and new polymers are introduced. As starting materials poly(1-hydroxy-1,3-propanediyl), poly(ethylene-co-vinyl alcohol), poly(vinyl alcohol), and poly(allyl alcohol) were used. The first three were cross-linked with a number of diisocyanate compounds. Network formation was performed at low concentrations in a good solvent. Mixing of the polymer solution and cross-linker appeared to be crucial for transparency. Poly(1-hydroxy-1,3-propanediyl), cross-linked with a slow reacting diisocyanate block, shows the most promising properties with respect to refractive index, transparency, tensile strength, and modulus. Poly(allyl alcohol) hydrogel was made by compression molding. The hydrogel was transparent and had a high refractive index and low modulus. It was concluded that hydrogels could be used as accommodating lens material.     

改良敷料联合早期康复锻炼在毒蛇咬伤患者中的应用

目的探讨改良换药法联合康复锻炼在提高毒蛇咬伤患者生活质量中的应用效果。方法选择2015年1月~2016年7月我院收治的120例蛇咬伤患者为研究对象,按随机数字表法分为常规组和改良组各60例,两组均采取清创排毒、扩创排毒、局部封闭、刺络放血法、外敷双黄蛇伤散(我院自制粉末状蛇药)、全身用药等综合治疗及常规蛇伤护理措施。在此基础上,常规组予带孔透明聚乙烯胶纸敷料包扎伤口,关节处用胶布固定;改良组予弹性防水透气防渗敷料包扎伤口,辅以弹力带适当外固定,治疗后3~7天指导患者每天进行关节康复运动。两组观察时间均为7天。结果治疗期间,两组患者均无明显不良反应。治疗后,改良组患者患肢疼痛消退时间、肿胀消退时间、肢体活动功能障碍、敷料评分、换药次数及住院时间均明显少于常规组,两组比较差异有统计学意义(t=3.599、3.583、2.703、2.178、3.394、6.384,均P0.05);改良组生活质量评分显著优于常规组,差异有统计学意义(t=6.642,P0.05)。结论采用改良敷料包扎蛇伤伤口并辅以弹力带外固定,具有稳固性强、美观、灵活的优点,便于患者关节活动,且可消肿止痛,减少换药次数,提高护理质量和效率;同时联合早期康复锻炼,有助于促进关节功能康复,改善患者生活质量,缩短住院时间,减轻患者经济负担,是一种安全、有效的康复锻炼方法,值得临床推广应用。     

Chiral separation of phospine‐containing α‐amino acid derivatives using two complementary cellulosic stationary phases in supercritical fluid chromatography

Enantiomeric separations of six amino‐acid derivatives have been studied using packed‐column supercritical fluid chromatography with two polysaccharide‐based enantioselective stationary phases: cellulose tris(3,5‐dimethylphenylcarbamate) and cellulose tris(3‐chloro‐4‐methylphenylcarbamate) (Lux Cellulose‐1 and ‐2). The effect of analyte structure on retention and separation was studied. Varied mobile phase compositions were investigated: alcohol modifier percentage was increased from 3 to 40% but smaller amounts were most effective in separating these compounds. Besides, ethanol was preferred to methanol or isopropanol as it proved to be a good compromise to achieve sufficient resolution in a reasonable analysis time. Moreover, a carbon dioxide‐ethanol mixture allows performing analyses in safe and green conditions. The effect of temperature at constant mobile phase composition was explored between 10 and 40°C. In most cases, increasing the temperature improved the chiral separation, up to an optimum temperature. The results are discussed in line with the structure variation of the racemic derivatives analyzed and the two columns are compared. The two columns were shown to provide complementary selectivities for the investigated solutes: whereas Lux 1 provided separation for five of the six racemates, Lux 2 could resolve the last racemic mixture. Finally, optimized conditions of separation are defined. Chirality, 2010. © 2009 Wiley‐Liss, Inc.     

两种不同方法制备的黑木耳多糖性质比较

本研究以黑木耳子实体为材料,对比了壳聚糖絮凝法制备的絮凝多糖HJD-1和传统水提醇沉法制备的醇沉多糖HJD-2的表观结构、α-葡萄糖甘酶抑制活性以及体外抑制肿瘤细胞增殖的活性。结果表明：（1）壳聚糖絮凝法制备粗多糖得率均值为4.76%,是醇沉法的2.17倍;醇沉法制备多糖的损失率为33.87%,是絮凝法的1.36倍;（2）絮凝多糖HJD-1和醇沉多糖HJD-2的表观评估及复溶性结果分析显示：絮凝多糖HJD-1为亮白色透明晶体,色泽均匀,颗粒规整;醇沉多糖HJD-2为棕褐色,颗粒状,有砂质感,前者相较后者的复溶性更好;（3）对α-葡萄糖甘酶抑制活性以及体外抗肿瘤能力结果分析表明：在相同浓度下,壳聚糖絮凝法制备黑木耳多糖对α-葡萄糖苷酶活性的抑制效果优于醇提法;絮凝多糖HJD-1对HepG2细胞的增殖抑制作用强于醇沉多糖HJD-2。     

Staining Paraffin Sections with Protargol 5. Chloral Hydrate Mixtures, with and Without Formamide, for Fixing Peripheral Nerves

A series of experiments was directed toward finding a means of improving fixation of mammalian glandular tissue and peripheral nerves with chloral hydrate. Specimens from cat, dog, rat, guinea pig, and man were fixed in solutions of 5-15% chloral hydrate in ethyl, methyl, and propyl alcohols, both pure and diluted with varying amounts of water. Modifiers were added, including acids, alkalies, alkaloids, amines, formamide, pyridine, piperidine, and formalin. The sectioned material was stained by the 2-hour method (AgNO₃-protargol) described previously (Davenport et al., 1939). The acidification of alcoholic chloral hydrate mixtures was deleterious to fixation but alkalinization was not. Among the modifiers, formamide was the one which showed definite improvement of fixation. A 10% solution of formamide alone in 50% ethyl alcohol gave good fixation and staining of peripheral nerve trunks, but addition of 5-7% chloral hydrate to this mixture improved staining. Treatment with 1% ammoniated alcohol after fixation and before embedding was of no value in section staining. Block stains were not tried.     

Spectroscopy analysis for simultaneous determination of lycopene and β-carotene in fungal biomass of Blakeslea trispora

Blakeslea trispora is a good alternative source for producing such carotenoids as lycopene and β-carotene. The objective of this research was to elaborate a method for the simultaneous determination of lycopene and β-carotene in Blakeslea trispora products using a usual UV-vis spectrophotometer. The standard solutions of the mixture of different concentrations of β-carotene and lycopene were measured with the UV-vis method and correlation formula for the extinction coefficients of 1% standard solution of lycopene in the solvent (hexane) and the ratios of the optical densities at the character peaks of 470 and 502 nm was elaborated. This gives a possibility to calculate the concentrations of lycopene and β-carotene in the mixture. The prediction quality of the UV-vis method was sufficient and the obtained results were very close to the ones, being measured with the HPLC technique. The proposed method can be used for both routine industrial work and academic research, providing the rapid analysis for simultaneous measurements of lycopene and β-carotene.