荧光素钠和考马斯亮蓝应用于小麦白粉病菌染色效果比较*

比较了荧光素钠和考马斯亮蓝应用于小麦白粉病菌染色的效果。荧光素钠法中样品处理只需20min.左右,具有直接、快速的特点;荧光指示剂对病菌分生孢子萌发及菌丝生长无抑制作用,主要沉集于活菌体的隔膜和细胞质部位,使病菌产生明显的亮绿荧光和清晰的细胞轮廓,亮绿荧光衰退期为7min.;借助荧光显微镜可以观察病菌在小麦叶表的发展过程,区别活菌体和失活菌体。考马斯亮蓝法包括传统的组织学染色步骤,经过改进后的样品处理过程需要40min.左右;染色后使寄主组织呈现淡蓝色,病菌菌体染成深蓝色;该方法可以观察病菌在小麦叶表和被侵染细胞内部发育形成的结构,包括孢子发育形成的初生芽管、附着胞芽管、成熟附着胞以及在寄主细胞内形成的初生吸器原体、成熟的指状体吸器和次生吸器。     

荧光素钠和考马斯亮蓝应用于小麦白粉病菌染色效果比较

比较了荧光素钠和考马斯亮蓝应用于小麦白粉菌染色的效果。荧光素钠法中样品处理只需20min左右,具有直接,快速的特点;荧光指示剂对病菌分生孢子萌发及菌丝生长无抑制作用,主要沉集于活菌体的隔膜和细胞质部位,使病菌产生明显的亮绿荧光和清晰的细胞轮廓,亮绿荧光衰退期为7min;借助荧光显微镜可以观察病菌在小麦叶表的发展过程,区别活菌体和失活菌体。考马斯亮蓝法包括传统的组织学染色步骤,经过改进后的样品处理过程需要40min左右;染后使寄主组织呈现淡蓝色,病菌菌体染成深蓝色;该方法可以观察病菌在小麦叶表和被侵染细胞内部发育形成的结构,包括孢子发育形成的初生芽管,附着胞芽管,成熟附着胞以及在寄主细胞内形成的初生吸器原体,成熟的指状体吸器和次生吸器。     

两种常用的免疫组化染色方法对NOS染色效果的比较

目的比较ABC法和SP法两种免疫组织化学染色方法的染色效果。方法通过免疫组化检测一氧化氮合酶的三个亚型的蛋白i NOS、e NOS和n NOS的表达,观察三种蛋白的染色结果。结果 SP法检测i NOS和e NOS的敏感性要明显强于ABC法;SP法蛋白染色结果明显优于ABC法。结论 SP法检测NOS优于ABC法。     

纤维蛋白的两种特殊染色效果比较

目的 比较分析显示纤维蛋白的两种特殊染色方法 ,旨在选择一种更适合于显示纤维蛋白的特殊染色方法。方法对富含纤维蛋白的石蜡包埋组织进行改良Masson染色、PTAH染色,光学显微镜下观察染色效果。结果 HE染色见纤维蛋白多数呈红染的细丝、相互连接成网状,也可相互融合。改良Masson染色显示纤维蛋白呈红色,PTAH染色显示纤维蛋白呈深蓝色。结论 改良Masson染色和PTAH染色均可用于显示纤维蛋白,但改良Masson染色时间短,结构清晰、对比明显,是观察纤维蛋白染色效果最佳的染色方法。     

几种染色剂对桔小实蝇产卵孔的染色效果

为寻找水果上桔小实蝇产卵情况的快速检测方法,本文测定了甲基蓝、龙胆紫、品红、曙红、藏红和刚果红等6种染色剂对水果上桔小实蝇Bactrocera dorsalis（Hendel）产卵孔的染色效果。结果表明染色剂对可疑水果进行染色,若有产卵孔,产卵孔可迅速被染色,染色率最高可达100％。筛选出甲基蓝、藏红和刚果红作为芒果、番石榴和夏橙的最适染色剂,各种染色剂在浓度为0．5％时染色效果较佳,同时对持续冷藏保存后的水果上的产卵孔仍然具有较高的染色率。     

两种切片方法检测肝糖原染色效果的比较

目的比较石蜡和冰冻两种不同切片用于检测牛蛙肝糖原的效果。方法采用石蜡和冰冻两种切片方法制作牛蛙肝脏切片,高碘酸希夫氏（periodicacid-Schiff＇s,PAS）染色法对肝糖原进行组织化学染色,光密度分析糖原含量。结果PAS染色显示肝糖原为紫红色或红色颗粒,冰冻切片中糖原颗粒明显大于石蜡切片,光密度分析显示,石蜡切片中糖原流失明显,与冰冻切片相比,糖原流失了约28％。结论两种切片均可用于糖原检测,冰冻切片制作环节较少、耗时短且染色过程中糖原不易流失。     

一种鉴定小鼠胚胎质量的双重荧光染色方法

尝试改进并建立一种较可靠的、适合小鼠胚胎质量参考鉴定的双重荧光染色(细胞计数)方法.与传统方法相比,本实验中抗羊脾细胞抗血清的最佳稀释度为1∶5,作用时间为30 min.豚鼠补体最佳稀释度为1∶5,两种分染染料H33342和PI的工作浓度均降低至5.3 μg/ml便可着色,染色时间延长至90 min,加大了染料工作液中柠檬酸钠的浓度,并在计数观察前增加压片这一步骤.染色过程中应注意避免血清物质对抗体补体的不良影响以及避光和快速的操作.双重荧光染色后小鼠胚胎内细胞团(ICM)细胞着色为蓝色,滋养层(TE)细胞着色为粉红色,数目清晰可辨.此方法可以作为一种简洁有效的小鼠胚胎质量鉴别参考方法.     

2种核酸染色方法的比较

目的:比较前染和后染等2种核酸染色方法对凝胶中DNA的染色效果,及其对染料染色灵敏度的影响,并验证新型核酸染色剂SYBRGreenⅠ能否代替传统染料溴化乙锭(EB)用于常规电泳中DNA的染色。方法:分别用SYBRGreenⅠ与EB采用2种不同的方法对凝胶中的DNA进行染色。结果:前染和后染的染色效果无显著差别,2种染料都能显示出10ng以下的DNA条带。结论:前染和后染方法的染色效果相当,染料SYBRGreenⅠ和EB均可用于这2种染色方法;新型染料SYBRGreenⅠ可代替强致癌性染料EB,用于常规凝胶中DNA的染色。     

果蝇唾腺染色体的几种染色方法比较

《生物学通报》编委 :您好。笔者对贵刊 2 0 0 2年第 37卷第 2期第 2 3页刊登的“用 Feuglen染色法制作果蝇唾腺染色体”一文感兴趣 ,但笔者认为文中几处不妥 ,比如 1)水浴温度波幅偏高 ;2 )染色时间不确定 ;3)试剂配方不全 ;4 )盐酸的浓度单位有误 ,mol?M?;5 )注意事项影响实验结果表述不清。特撰写下文与各位读者商榷。双翅类昆虫如黑腹果蝇 (Drosophila melanogaster)的唾腺染色体 (Salivary chromosome)比普通染色体大的多 ,处于体细胞同源染色体的配对状态 ,是由于唾腺染色体经过多次复制而并不分开形成的 ,大约有 10 0 0～4 0 0 0根…     

两种核酸染料染色效果的比较研究

用前染和后染两种不同的染色方法,研究比较SYBRGreenI和溴化乙锭(EB)两种核酸染料对凝胶中DNA的染色效果和灵敏度,及SYBRGreenI取代EB用于常规凝胶中核酸染色的可能性。结果表明,用前染法染色SYBRGreenI对琼脂糖凝胶中的核酸染色效果与EB相当;用后染法染色前者要优于后者,可显示5ng以下的DNA条带,在完全相同的操作条件下,其染色DNA条带背景清晰,灵敏度较高。因此,无致突变性新型染料SYBRGreenI可替代强致突变性染料EB用于检测凝胶中DNA片段大小、含量等,从而减少由于使用EB带来的环境污染和人体健康危害。     

石蜡组织荧光原位杂交关键实验步骤探讨

目的：探讨石蜡组织荧光原位杂交（FISH）技术中关键实验步骤的最佳条件,以期提高石蜡切片FISH阳性细胞检出率和实验成功率。方法：在前期标本处理方面,进行双蒸水、0．5×SSC溶液、0．1％硫代亚硫酸钠溶液煮沸对比;在蛋白酶消化方面,设置胃蛋白酶和蛋白酶K两种消化方法,并在37℃条件下设置时间的梯度变化,比较其消化效果;对石蜡切片的变性设置温度梯度,比较杂交检出率;比较DAPI复染时不同浓度对单色、双色FISH结果的影响;应用抗淬灭剂后不同保存时间的对比。结果：采用0．5×SSC溶液煮沸15min,用200μg／mL蛋白酶K在37℃、6～10min条件下消化,可以取得较好的FISH效果;变性温度为81℃时检出率更高,DAPI复染浓度为1000ng／mL时针对单色FISH较合适,而浓度为500、150ng／mL时针对双色／多色FISH有较好的效果。结论：FISH条件经过对比得到优化,对石蜡组织FISH实验具有一定的指导意义。     

玉米rRNA基因的组织和表达模式

玉米（Zea mays）只有1对45S rDNA位点并在分裂期染色体形成次缢痕,是研究植物细胞rRNA基因组织和表达模式的简单模型。采用荧光原位杂交（fluorescence in situ hybridization,FISH）、CPD（PI与DAPI组合）染色和银染技术,分析了玉米根尖分生细胞rRNA基因的组织和表达模式。45S rDNA探针在所有间期细胞核中显示2种杂交信号：荧光强烈地位于核仁周边的纽,而相对较弱地分布于核仁内的点。在部分细胞中可观察到点与纽相连或从纽发出;点的数目越多,纽变得越小;点的数目多少与细胞的活性呈正相关。研究结果表明,纽代表了处于凝缩状态的非活性的rDNA染色质,纽解凝缩形成的点是rRNA基因活跃转录的细胞学表现;不同阶段间期核的点的数目变化反映了被活化的rRNA基因数目不同。间期和前期细胞的CPD染色和相继的银染结果显示,大部分rDNA染色质没有参与核仁的形成。rDNA FISH显示,同一间期细胞的2个同源rDNA位点的表达水平存在差异,同源染色体次缢痕的长度差异以及Ag-NOR和银染核仁的异态性进一步证实了这种差异的存在。FISH结果显示,早中期细胞的rDNA染色质相对解凝缩,银染在所有早中期细胞和部分中期细胞显示了明显的核仁,表明玉米的rRNA基因在有丝分裂早中期有较活跃的转录,其转录在晚中期才停止。     

Abnormal chromosome 8 copy number in cytological smears from breast carcinomas detected by means of fluorescence in situ hybridization (FISH)

Numerical change in chromosome 8 is an acquired abnormality associated with high clinical stage and may be involved in the conversion of carcinoma in situ in the breast to invasive carcinoma. Fine needle aspiration smears from 53 cases of breast carcinoma were hybridized with centromeric probes for chromosome 8 and the X chromosome. Thirty-eight cases revealed chromosome 8 copy gain. Of the 45 grade II and III tumours, 28 showed polysomy (>3 signals) and six showed trisomy. Of the eight grade I tumours, four were trisomic, none were polysomic. There were only two cases of chromosome 8 copy loss (one each of grade I and III). X chromosome polysomy was also a frequent finding although the signal counts were similar to those for chromosome 8 in only a few cases. Chromosome 8 polysomy occurs frequently in breast carcinoma and high copy number (>3) is associated with high malignancy grade.     

荧光原位杂交技术分析栽培种甘薯(Ipomoea batatas cv.XushuNo.18)染色体

为了解栽培种甘薯(徐薯18,Ipomoea batatas cv.XushuNo.18)的染色体结构,文章利用45SrDNA荧光原位杂交、自身基因组荧光原位杂交和银染技术对栽培种甘薯进行分子细胞遗传学研究。银染结果显示,徐薯18间期核有6对、8对和9对银染点;45SrDNA荧光原位杂交结果显示,徐薯18染色体上有8对或9对强弱不一的45SrDNA信号;自身基因组荧光原位杂交结果表明,所有染色体的全长分布强烈而密集的杂交信号,着丝粒区、近着丝粒区和端粒区有增强的信号带。     

45S rDNA基因的不稳定性及其与转录的相关性

45S rDNA基因由串联重复序列构成,是遗传不稳定性的热点区域,易于发生DNA断裂和重组。以Hela和CHO细胞系为研究对象,运用荧光原位杂交技术检测有丝分裂不同时期的45S rDNA基因的不稳定性表型。结果表明,位点特异性的染色体浓缩失败是其在中期染色体上不稳定性的主要表型。具有这种表型的染色体在后期可能会出现落后或粘连现象,甚至有可能引发断裂,形成卫星核。同时,免疫荧光双染色技术检测表明DNA双链断裂的标记蛋白(γH2AX)和RNA聚合酶I的上游结合因子(UBF)在有丝分裂的不同时期都存在共定位现象。该结果为探讨45S rDNA基因的不稳定性与转录的关系提供了直观的细胞学证据。     

A Review of Fluorescence in Situ Hybridization (FISH): Current Status and Future Prospects

Fluorescence in situ hybridization (FISH) is a powerful technique for detecting DNA or RNA sequences in cells, tissues and tumors. This molecular cytogenetic technique enables the localization of specific DNA sequences within interphase chromatin and metaphase chromosomes and the identification of both structural and numerical chromosome changes. FISH is quickly becoming one of the most extensively used cytochemical staining techniques owing to its sensitivity and versatility, and with the improvement of current technology and cost effectiveness, its use will surely continue to expand. Here we review the wide variety of current applications and future prospects of FISH technology.     

A Review of Fluorescence in Situ Hybridization (FISH): Current Status and Future Prospects

Fluorescence in situ hybridization (FISH) is a powerful technique for detecting DNA or RNA sequences in cells, tissues and tumors. This molecular cytogenetic technique enables the localization of specific DNA sequences within interphase chromatin and metaphase chromosomes and the identification of both structural and numerical chromosome changes. FISH is quickly becoming one of the most extensively used cytochemical staining techniques owing to its sensitivity and versatility, and with the improvement of current technology and cost effectiveness, its use will surely continue to expand. Here we review the wide variety of current applications and future prospects of FISH technology.     

Optimization of PRINS and C-PRINS for detection of telomeric sequences in Vicia faba

Primed in situ labelling (PRINS) of nucleic acids was developed as an alternative to traditionally used fluorescence in situ hybridization (FISH). Compared to FISH, PRINS is faster and does not require preparation of labelled probes. Nevertheless, the number of applications for physical mapping of DNA sequences on plant chromosomes remains low. This is due to the fact that there are a number of factors which influence the specificity and sensitivity of the reaction. The purpose of this work was to analyse the effect of some of them, including the age of slides, type of Taq DNA polymerase, number and concentration of primers, the presence and concentration of bovine serum albumine and MgCl₂ in the reaction mixture. Furthermore, the effect of various pre-treatments on signal intensity and non-specific fluorescence was studied. A consensus Arabidopsis-type telomeric sequence and Vicia faba mitotic chromosomes were used as a model system. We have found that the age of slides was critical and that under optimal conditions it was possible to achieve relatively high signal to noise ratio. This revised version was published online in July 2006 with corrections to the Cover Date.     

The potential of flow cytometry in the study of Bacillus cereus

Flow cytometry (FCM) is a rapid method allowing the acquisition of multiparametric data from thousands of individual cells within a sample. As well as measuring the intrinsic light scattering properties of cells, a plethora of fluorescent dyes may be employed to yield information on macromolecule content, surface antigens present or physiological status. Despite FCM's indispensability within other fields e.g. immunology, it is underutilized within microbiological research. In this review, a strong case is presented for the potential of FCM in the study of Gram-positive spore-former, Bacillus cereus . Previous reports where FCM was successfully used in the study of B. cereus are reviewed along with relevant studies involving other members of the genus. Under headings reflecting common research themes associated with B. cereus , specific instances where FCM has generated novel data, providing a unique insight into the organism, are discussed. Further applications are posited, based on the authors' own research with FCM and B. cereus and work extant in the broader field of microbial cytometry. The authors conclude that, while the expense of equipment and reagents is an undeniable disadvantage, FCM is a technique capable of generating significantly novel data and allows the design and execution of experiments that are not possible with any other technique.     

Effect of Explosion on the Crystalline Polymorphism of Chitin and Chitosan

For identification of how explosion increases the reactivity of chitin and chitosan, changes in the crystalline polymorphism of these polysaccharides were studied by X-ray diffraction measurements. The α-chitin form of chitin did not change after being exploded, but an X-ray diagram of chitosan showed a hydrated crystal of low crystallinity before the explosion, and increased crystallinity of the hydrated form plus a small amount of an anhydrous crystal after the explosion. The improvement of accessibility to both polysaccharides caused by the explosion seemed not to arise from changes in their crystalline polymorphism or crystallinity