常规石蜡切片方法的改良

针对传统石蜡切片方法中的缺陷,对制片方法进行了相应的改良。总结了切片制作过程中可能存在的问题以及处理对策;提出了一些能缩短实验周期,解决实验有毒物质二甲苯污染的方案。结合教学实践发现改良方案有助于提高石蜡切片的质量。     

石蜡切片贴片法的改良

传统的石蜡切片贴片操作费时,而且效果不太理想。作者经过多次摸索试验,对其进行了改良,取得了良好的效果。     

长松萝多糖的研究

长松萝经三氯甲烷抽提后风干,用热水提取,乙醇沉淀,经微晶纤维素柱层析纯化,得白色粉末状多糖USL。USL经Sephadex G-150柱层析证明为一组均匀多糖,其糖的含量为89.52％。经气相色谱检定,由阿拉伯糖(Ara)、木糖(Xy1)、甘露糖(Man)、葡萄糖(Glc)聚合而成,其克分子比为0.31:0.05:1.00:18.10。经Sephadex G-200柱层析测定,平均分子量为30×10~4,经高碘酸氧化,Smith降解,有甲酸、丙三醇、赤藓醇生成。红外光谱在896cm-~1处有吸收,证明USL多糖结构主要以β(1→4)、β(1→6)甙键连接而成的杂多糖。     

石蜡切片刀的改进研究

以手术刀片代替传统的切片刀,通过肝、肾、脾、肺、心肌、气管等器官所作的对比性切片试验表明,该切片刀不仅达到了原有切片刀的性能,还能做1—3微米半薄切片。染色后在10×10、10×40显微镜下,整个切片完整无损,厚度均匀,各类细胞结构清晰易辨。该切片刀不需磨鐾,省时省力,操作简便,经济实用。特别适合基层实验室的组织及病理学的教学科研工作。     

新疆松萝属地衣的研究

松萝属地衣在世界范围内是研究比较深入。本文概述了松萝属地衣在新疆的研究历史及现状,阐述了新疆松萝属地衣的物种鉴定、所含地衣物质及其药用价值,以及松萝酸和松萝多糖等的相关研究进展,并对松萝属地衣研究的发展趋势作了展望。     

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。     

动物毛发石蜡切片的制作

目的:探讨哺乳动物针毛横断面的快速简易的制作方法,从毛发横断面上观察毛发微观结构。方法:取新疆部分哺乳动物(马鹿塔里木亚种Cervus elaphus yarkandensis,藏羚羊Pantholops hodgsoni,野双峰驼Camelus bactrianus,狍子Capreolus capreolus)的毛发,先将其清洗然后经浸蜡、包埋、切片、染色、封片等一系列过程的研究,得发毛横断面。结果:各动物毛发微观结构差异显著:马鹿塔里木亚种,毛皮质极薄,毛髓质占大多数;藏羚羊,由若干多边行空囊组成,内部中空;野双峰驼,毛皮质与毛髓质所占比例相当;狍子,毛皮质约占2/3,毛髓质约占1/3。结论:为利用哺乳动物毛发进行种属鉴定奠定一定科学理论基础。     

兔卵巢组织石蜡切片技术的改良

针对常规石蜡切片方法步骤繁多,耗时长,所用的二甲苯试剂对人体健康有较大毒害性的缺陷,探讨提高制作兔卵巢组织石蜡切片质量的方法。试验从脱水和透明环节进行技术改良,用正丁醇与无水乙醇混合剂取代常规的脱水剂和透明剂。该混合剂具有良好的脱水和透明作用。实验结果表明：通过该技术改良,镜下观察组织结构清晰可辨、颜色鲜艳、对比度好、层次分明,组织切片效果良好。试验改良技术的工艺简单,具有质优、省时、快捷和实用性较好等优点,有推广应用价值。     

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。     

水稻籽粒胚乳的石蜡切片方法改良及其结构发育观察

以杂交水稻保持系‘岗46B’授粉后不同天数的籽粒为材料,利用PAS反应原理,通过对传统石蜡切片技术中材料固定、染色、脱水、浸蜡等环节的改进,即采用FAA和FPA相结合方式固定材料,先用热配法席夫(Schiff)试剂进行整染,再以稀释和氨水蓝化后的埃利希氏(Ehrlich)苏木精复染,中间以70%、83%、95%质量分数梯度乙醇进行脱水和复水处理,渐进式浸蜡与把握时间等实验环节,对水稻胚乳结构发育进行观察研究,结果授粉后第2~10天籽粒胚乳的石蜡切片完整,且染色分明,结构显示完整清晰。观察发现,随着授粉后天数推移,胚乳内部逐渐由游离胚乳核期向细胞化过渡,细胞层数增加并趋向充满子房,淀粉粒开始累积,胚乳细胞进一步增大。授粉后第5~10天是胚乳细胞淀粉粒从开始到大量累积的重要时期,同时胚乳细胞向无核化发展,不同部位胚乳细胞和淀粉粒的排列分布也逐渐表现出一定的差异。表明改良后的石蜡切片方法适合于授粉后第0~10天的水稻胚乳结构发育研究。     

Comparative Ploidy Studies Using Cytological and Paraffin Section Preparations

The ploidy profiles of benign and malignant tumours can be obtained using image analysis. However, the results of ploidy studies have varied according to the type of specimen used. We compared the ploidy profiles of paraffin embedded thin sections, cytospin preparations of disaggregated cells, and cytological smears from the same specimen as defined by image analysis. Ten benign breast lesions, 10 breast carcinomas and 10 malignant melanomas were investigated in this way. Preparations stained by the Feulgen technique were examined using the MD20 video image analysis densitometry system. Ploidy profiles were obtained by measuring the integrated optical density of at least 200 nuclei. By paying proper attention to the quality of fixation and presence or absence of cytoplasm around cells, comparable results were found for all preparations in each case. We therefore conclude that if careful attention is paid to the technical quality of the material, reliable ploidy results can be obtained by image analysis.     

Influences of Nitrogen Sources on Usnic Acid Production in a Cultured Mycobiont of the Lichen Usnea hirta (L.) Wigg.

Effects of the nitrogen sources in the medium for the production of secondary metabolites in lichens were examined. The usnic acid production by a mycobiont of the lichen Usnea hirta was higher in the liquid medium containing ammonium and nitrate ions than in those containing amino acids.     

两栖类皮肤石蜡切片样本量探讨——以峨眉髭蟾毛细血管丰富度为例

石蜡切片是量化两栖动物各种皮肤特征的常用技术,但在两栖动物中缺乏取样标准。血管密度等无法直接度量的特性可能需要较大的样本量,探讨这类特征的取样标准有利于实践工作。本研究对雄性峨眉髭蟾Leptobrachium boringii表皮下毛细血管丰富度进行量化,并通过重抽样分析该特征所需样本量。实验获得2只个体10个部位数据,每个部位模拟29种抽样。结果显示,体型较小个体多数部位需接近20条组织或更高的样本量,体型较大个体多数部位所需样本量接近15条或更高。建议在两栖类皮肤石蜡切片工作中对全部有效切片进行制片,以满足不同特征对样本量的要求。表皮下毛细血管丰富度与所需样本量呈负相关关系,可能缘于高密度部位的血管分布更均匀。如在其他两栖动物类群中亦观察到该相关性,则有助于在量化该特征时合理选取样本量。     

Observation on Flower Bud Differentiation of Crape Myrtle in Red Soil Environment

Flower bud differentiation is a key component of plant blooming biology and understanding how it works is vital for flowering regulation and plant genetic breeding, increasing the number and quality of flowering. Red soil is the most widely covered soil type in the world, and it is also the most suitable soil type for crape myrtle planting. The flower buds of crape myrtle (Lagerstroemia indica) planted in red soil were employed as experimental materials in this study, and the distinct periods of differentiation were identified using stereomicroscopy and paraffin sectioning. We optimized the steps of dehydration, transparency, embedding, sectioning and staining when employing paraffin sections. When seen under a microscope, this optimization can make the cell structure of paraffin sections obvious, the tissue structure complete, and the staining clear and natural. The flower bud differentiation process is divided into 7 periods based on anatomical observations of the external morphology and internal structure during flower bud differentiation: undifferentiated period, start of differentiation period, inflorescence differentiation period, calyx differentiation period, petal differentiation period, stamen differentiation period, and pistil differentiation period. The differentiation time is concentrated from the end of May to mid-June. Crape myrtle flower bud differentiation is a complicated process, and the specific regulatory mechanism and affecting elements need to be investigated further.     

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。     

Antioxidant and antibacterial activities of lichen <Emphasis Type="Italic">Usnea ghattensis</Emphasis><Emphasis Type="Italic">in vitro</Emphasis>

Various solvent extracts of the lichen Usnea ghattensis showed good antioxidant activity. A methanol extract prevented lipid peroxidation by 87% followed by 65% in Trolox at 20 μg/ml. It also showed superoxide anion scavenging activity and free radical scavenging activity 56% and 73%, respectively. The known antioxidants butylated hydroxytoluene (BHT), butylated hydroxyanisol (BHA) and quercetin at similar concentrations showed superoxide anion scavenging activity of 68, 59 and 47% and free radical scavenging activity 83, 77 and 69%, respectively. In addition, these extracts were inhibitory against Bacillus licheniformis, Bacillus megaterium, Bacillus subtilis and Staphylococcus aureus with MIC values of 5–10 μg/ml.Received after revisions 10 May 2005     

高寒冰缘区15种藓类植物茎的形态结构研究

采用石蜡切片方法和扫描电镜方法,对高寒冰缘区生长的15种藓类植物茎的结构进行解剖学观察分析,为高寒冰缘区藓类植物在极端环境下的生存机理及其生态应用提供依据。结果表明:高寒冰缘区的15种藓类植物的茎分为表皮、皮部、中轴三部分。茎表皮细胞多为一层,长短不一,细胞壁表面粗糙,表面纹饰多为鳞片状、颗粒状、浅窝点状,表皮细胞的细胞壁加厚程度不同;皮部所占面积最大,大部分有内、外皮部的分化,细胞壁由外向内逐渐变薄,细胞由小到大又到小的整齐或镶嵌排列;中轴分化但其细胞数多少不一,细胞壁多具角隅加厚。研究认为,高寒冰缘区藓类植物具有典型的耐寒、耐旱结构特征,这些结构特征能够增强藓类植物的支撑和传导水分的作用,以抵御长期寒冷、干旱、多强风等恶劣环境;其中细胞的形状、数目、细胞表面的纹饰等微形态结构特征也具有潜在的分类学意义。     

小花山桃草营养器官解剖结构及其生态适应性研究

通过石蜡切片法对外来入侵植物小花山桃草进行解剖学方面的研究,旨在揭示其入侵和蔓延的结构基础。结果表明：小花山桃草根的次生结构中次生木质部所占比例较大,约占整个横切面的2/3,导管数量多,平均达138.25个,管腔大,管径为85.37 μm;根和茎的木栓层均较发达,由6～7层扁平细胞组成;根和茎的次生韧皮部中存在大量含针晶细胞;小花山桃草的叶具典型的旱生植物叶片的结构特征：表皮为复表皮,上下表皮均有气孔分布,上表皮气孔密度为180 mm^-2,下表皮气孔密度为266 mm^-2;栅栏组织为双栅型,近轴面栅栏组织细胞2～3层,排列紧密而整齐,含叶绿体较多;叶片主脉木质部发达,由多列导管组成。上述特征说明小花山桃草的解剖结构对干旱生境有较强的适应性。