植物小孢子母细胞减数分裂过程中胼胝质染色的新方法

利用改良苯酚品红-苯胺蓝压片法,观察小孢子母细胞减数分裂过程中胼胝质的动态变化。使用该方法简便、快速且省时,获得的照片颜色鲜艳,细胞质呈红色,染色体为深红色,胼胝质呈黄绿色荧光,对比明显,有三维效果。单用改良苯酚品红染液对新鲜材料进行压片,在蓝光激发下,细胞质与染色体呈红色荧光,染色体清晰。实验结果表明,改良苯酚品红染液可作为荧光染料代替DAPI及H33258等昂贵的核染料,从而降低实验成本。     

橘小实蝇染色体研究与相关技术探讨

以橘小实蝇Bactroceradorsalis(Hendel)性腺和唾液腺为材料,采取改良苯酚品红染色后压片法,制备染色体标本。对橘小实蝇进行染色体组型研究和唾液腺染色体形态学的观察。结果表明:橘小实蝇成虫性腺染色体数目为2n=12条,染色体长度基本呈连续性变化,其性染色体属于XX/XY型。幼虫唾液腺染色体包含5条较长的多线染色体,与成虫常染色体相对应。实验显示苯酚品红染色后压片为制备染色体标本简单有效的方法。     

用改良苯酚品红染色液替代醋酸洋红染色液的研究

以往在动物遗传学实验“果蝇唾液腺染色体标本的制作和观察”中,采用醋酸洋红染色液对染色体染色。本文对用改良苯酚品红染色液替代醋酸洋红染色液对果蝇唾液腺染色体染色的问题进行了研究。结果表明,改良苯酚品红染色液对果蝇唾液腺染色体的染色效果与醋酸染洋红染色液的染色效果是相同的。而且用改良苯酚品红染色液还人提高工效,简易节约的优点。因此认为,在对果蝇唾液腺染色体染色中,用改良苯酚品红染色液替代酸酸洋红染色液     

果蝇唾腺染色体制片方法的改进

对果蝇唾腺染色体制片方法进行了改进．结果表明：改良苯酚品红染色液配好15d后使用,浓度为6％时,染色5～8min可以获得染色体横纹和背景清晰的图像;通过改变脂肪剥离的顺序即染色后再去除脂肪,压片效果明显提高。     

甘薯减数分裂观察方法

甘薯的染色体很小,而且数量较多(2n=90),同时又是六倍体植物,减数分裂行为较为复杂,所以,其减数分裂的细胞学观察也很困难。国内很少研究,国外报道一般还是采用传统的醋酸洋红压片法。但染色体分散不好,细胞质着色深,很难获得理想的分裂相,作减数分裂遗传分析时,往往还得借助人工绘图加以说明。本试验对醋酸洋红压片技术略加改良,采用压片(醋酸洋红)—简易水解(浓盐酸:纯酒精=1:1)—压片(石碳酸品红)流     

人体细胞巴氏小体的观察

本文介绍一种简便、成功率高的人体细胞巴氏小体染色方法,具体方法如下：１染液采用改良苯酚品红染液。母液Ａ：３ｇ碱性品红,溶于１００ｍｌ７０％酒精中（可长期保存）。母液Ｂ：取１０ｍｌ母液Ａ,加入９０ｍＬ５％苯酚水溶液。取４５ｍｌ母液Ｂ,加入６ｍｌ冰醋酸和...     

桔梗花粉母细胞减数分裂及雄性败育的细胞生理学研究

以桔梗不育系PA及其保持系PB为试验材料,采用石蜡切片和改良苯酚品红染色压片法对花粉母细胞减数分裂和雄配子体发育过程进行比较,探讨桔梗雄性不育系小孢子形成过程和败育发生的细胞生理学机理。结果表明:(1)桔梗保持系PB花粉母细胞减数分裂的细胞质分裂为同时型,同一花药减数分裂较同步;在中期Ⅰ和中期Ⅱ,少数细胞中可见赤道板外染色体;四分体以四面体为主,成熟花粉粒为二核花粉。(2)不育系PA花粉母细胞减数分裂后期Ⅰ开始出现异常,表现为细胞质形态改变,末期Ⅱ之后细胞质不能分裂,形成异常四分体,胼胝质壁不能溶解,四分体难以释放出游离小孢子而被降解,导致败育。(3)在发育过程中,桔梗不育系花蕾游离脯氨酸、可溶性蛋白含量低于保持系,而SOD活性、丙二醛含量均高于保持系。(4)桔梗不育系PA及其保持系PB花粉母细胞减数分裂和雄配子体发育过程存在明显差异,桔梗雄性败育过程大体可分为4个阶段,即后期Ⅰ细胞质异常、末期Ⅱ之后细胞质不能分裂、四分体难以释放游离小孢子、四分体被降解仅残留碎片。研究认为,桔梗不育花蕾(开花前)生长发育过程中,体内活性氧代谢紊乱、丙二醛积累及游离脯氨酸等"物质代谢损亏"可能是引起桔梗雄性败育的原因。     

对DNA与RNA区分染色实验的改进

核酸是主要的遗传物质 ,核酸的主要成分在细胞核中是 DNA,在细胞质与核仁中的是 RNA。由于 DNA与RNA在化学组成与分子结构上存在一定差别 ,因而对不同染料有不同的染色反应 ,可以根据这一原理来定性鉴定细胞中 DNA与 RNA的存在与分布。实验所采用染料为甲基绿 -焦宁 (Netyl- Green-Pyronin)染液 ,其中染液中的甲基绿能使细胞核中的DNA呈现绿色 ,而焦宁则能把细胞质与核仁中的 RNA染成红色。因此可以根据细胞中不同部位呈现颜色的不同来进行定性鉴定。1　材料与方法为了使实验方法快速简易而效果清晰准确 ,我们从染液的配方、材料…     

不同浓度的苯酚品红对果蝇唾腺染色体的染色效果

以不同浓度的苯酚品红为染色液,观察其对果蝇唾腺染色体的染色效果,结果表明,8％－12％苯酚品红为染色液,可获得背景清楚,染色体横纹清晰的图像。     

九月份生物各学科实验准备

植物学(实验一和实验二) 材料用具:显微镜(低倍镜)、小块紫色洋葱鳞茎(若无紫色洋葱时,还需准备碘液染色)、内盛清水的小烧杯、吸管、镊子、刀片、牙签(代替解剖针)、载玻片、盖玻片、小片吸水纸、绘图纸,番茄、红色柿子椒各一个(演示用)。示范镜的准备:1.示成熟的番茄果肉细胞:用牙签挑取果肉细胞制成装片观察。2.示辣椒表皮细胞:将柿子椒用小刀切成小块,清除掉果肉制成无色表皮细胞,观察胞间连丝,如用卡宝染液(改良苯酚品红染色液)染色效果更好。卡宝染色液配制方法如下: ①取3克碱性品红,溶于100m170％酒精中,配成母液     

A new fluorescent staining method for callose of microspore mother cells during meiosis

To observe the dynamic behavior of callose of microspore mother cells during meiosis, we developed a convenient, rapid and efficient staining method using an improved carbol fuchsin/aniline blue solution. The stained microspore mother cells during meiosis showed yellowish green callose, red cytoplasm and dark red chromosomes when excited with blue light, which produced a contrasting image with a three-dimensional effect. When stained with only improved carbol fuchsin solution, the cells had red cytoplasm and chromosomes when excited with green light. The improved carbol fuchsin solution can be used to replace other more expensive DNA-specific dyes, such as DAPI and H33258, to reduce experimental costs.     

A new fluorescent staining method for callose of microspore mother cells during meiosis

To observe the dynamic behavior of callose of microspore mother cells during meiosis, we developed a convenient, rapid and efficient staining method using an improved carbol fuchsin/aniline blue solution. The stained microspore mother cells during meiosis showed yellowish green callose, red cytoplasm and dark red chromosomes when excited with blue light, which produced a contrasting image with a three-dimensional effect. When stained with only improved carbol fuchsin solution, the cells had red cytoplasm and chromosomes when excited with green light. The improved carbol fuchsin solution can be used to replace other more expensive DNA-specific dyes, such as DAPI and H33258, to reduce experimental costs.     

Demonstration of axial elements of sex vesicles of spermatocytes using Coomassie brilliant blue

The axial element of sex chromosomes in the sex vesicle of rat and mouse spermatocytes has been visualized under the light microscope by the dye Coomassie brilliant blue (CBB). After staining in the CBB solution for 3-10 minutes, the axial elements appeared as darkly stained threads in the sex vesicles, whereas in controls stained with Giemsa or carbol fuchsin, the sex vesicles were usually uniformly stained. The axial elements are best seen when chromosome preparations were made by the flame drying technique. In rat spermatocytes the staining quality could be further improved by a brief treatment with trypsin solution (0.025%). The CBB staining procedure is simple and easily controllable. The results suggest that the CBB stained material is protein in nature and is more resistant to trypsin digestion than other nuclear proteins.     

Demonstration of Axial Elements of Sex Vesicles of Spermatocytes Using Coomassie Brilliant Blue

The axial element of sex chromosomes in the sex vesicle of rat and mouse spermatocytes has been visualized under the light microscope by the dye Coomassie brilliant blue (CBB). After staining in the CBB solution for 3-10 minutes, the axial elements appeared as darkly stained threads in the sex vesicles, whereas in controls stained with Giemsa or carbol fuchsin, the sex vesicles were usually uniformly stained. The axial elements are best seen when chromosome preparations were made by the flame drying technique. In rat spermatocytes the staining quality could be further improved by a brief treatment with trypsin solution (0.025%).

The CBB staining procedure is simple and easily controllable. The results suggest that the CBB stained material is protein in nature and is more resistant to trypsin digestion than other nuclear proteins.     

The Germicidal Effect of Staining Solutions

Gentian violet, crystal violet and carbol fuchsin applied to cover slip preparations for one minute will destroy the majority of non-spore-forming bacteria and yeasts, tho they can not be relied upon to do this consistently and in all cases.

The Gram staining procedure is more effective and non-spore-formers were never found to survive this process.

Methylene blue stains exert very little if any germicidal power and most organisms survived them readily. India ink was totally ineffective.

Several species of yeasts and yeast-like molds were killed in every instance by the Gram stain, gentian violet, crystal violet and carbol fuchsin, but survived both Loeffler's methylene blue and a plain aqueous solution of methylene blue.     

The Germicidal Effect of Staining Solutions

Gentian violet, crystal violet and carbol fuchsin applied to cover slip preparations for one minute will destroy the majority of non-spore-forming bacteria and yeasts, tho they can not be relied upon to do this consistently and in all cases.

The Gram staining procedure is more effective and non-spore-formers were never found to survive this process.

Methylene blue stains exert very little if any germicidal power and most organisms survived them readily. India ink was totally ineffective.

Several species of yeasts and yeast-like molds were killed in every instance by the Gram stain, gentian violet, crystal violet and carbol fuchsin, but survived both Loeffler's methylene blue and a plain aqueous solution of methylene blue.     

A Simple Staining Method for the Analysis of Meiotic Metaphase II Divisions in the Male Mouse

During an investigation into the effects of X rays on meiosis in the male mouse (Szemere and Chandley 1975) a staining technique was required that would enable us to make an accurate analysis of dyads² at metaphase II. Not only were we interested in analysing chromosomal aberrations at this stage, but we also wished to identify with confidence the X and Y chromosomes, and to establish accurate counts of dyad numbers. Conventional staining with carbol fuchsin (Can and Walker 1961) provided adequate means for recognizing sex chromosomes, but centromere positions could not be identified and little morphological detail of autosomal dyads could be discerned. Staining by the BSG barium hydroxide/saline/Giemsa technique (Sumner 1972) as modified for use on meiotic cells of the mouse (Chandley and Fletcher 1973) gave excellent staining of centric heterochromatin, but dyad arms were often pale and indistinct. Other centromere staining methods for murine meiotic cells (Hsu, Cooper, Mace and Brinkley 1971, Polani 1972), gave unsatisfactory results in our hands. By combining carbol fuchsin staining with the BSG centromere staining technique, we have been able to produce a simple and quick technique which gives excellent staining of centromeres, easy identification of X and Y chromosomes and good staining of dyad arms at metaphase II. The technique has also been applied successfully to other meiotic stages of the mouse and to human somatic metaphase chromosomes.     

A Safe Spore Stain for Class Use

In looking for an efficient method of pre-treating bacterial spores for staining with carbol fuchsin, the author found that the ammonia in Lagerberg's formula and the chromic acid in Moeller's formula were both fairly effective in increasing permeability. Combining the two proved highly successful; and the following method has been used for four years by classes of beginners in the study of spore-formers isolated from soil by the students.

Cover the heat-fixed smear with five percent chromic acid and after thirty seconds add twice the amount of concentrated ammonia to the acid. After two minutes rinse and steam with carbol fuchsin for two or three minutes. Rinse, destain with one percent sulphuric acid for fifteen to thirty seconds, rinse again and flood slide with tap water. Add to this a few drops of Loeffler's methylene blue and allow to counter-stain for ten to thirty seconds. Rinse, blot, dry and examine. If instructions are followed the results are always good.     

A Simple Staining Method for the Analysis of Meiotic Metaphase II Divisions in the Male Mouse

During an investigation into the effects of X rays on meiosis in the male mouse (Szemere and Chandley 1975) a staining technique was required that would enable us to make an accurate analysis of dyads² at metaphase II. Not only were we interested in analysing chromosomal aberrations at this stage, but we also wished to identify with confidence the X and Y chromosomes, and to establish accurate counts of dyad numbers. Conventional staining with carbol fuchsin (Can and Walker 1961) provided adequate means for recognizing sex chromosomes, but centromere positions could not be identified and little morphological detail of autosomal dyads could be discerned. Staining by the BSG barium hydroxide/saline/Giemsa technique (Sumner 1972) as modified for use on meiotic cells of the mouse (Chandley and Fletcher 1973) gave excellent staining of centric heterochromatin, but dyad arms were often pale and indistinct. Other centromere staining methods for murine meiotic cells (Hsu, Cooper, Mace and Brinkley 1971, Polani 1972), gave unsatisfactory results in our hands. By combining carbol fuchsin staining with the BSG centromere staining technique, we have been able to produce a simple and quick technique which gives excellent staining of centromeres, easy identification of X and Y chromosomes and good staining of dyad arms at metaphase II. The technique has also been applied successfully to other meiotic stages of the mouse and to human somatic metaphase chromosomes.