一种改良的鱼类染色体富集制片方法

运用淋巴细胞分离液处理黄鳝肾脏细胞悬浮液后,成功地分离了其中红细胞和淋巴细胞。以富集的淋巴细胞进行染色体制片,可避免有核红细胞的影响,获得高分裂指数的黄鳝染色体标本。该方法简便、结果稳定,亦适用于其他鱼类、两栖类和鸟类的染色体制片。 Abstract：After treated by the lymphocyte separation medium, the erythrocytes and lymphocytes were separated successfully in the kidney cells suspension of rice field eel. Using the enriched lymphocytes to prepare chromosome, we get the high split-index chromosome preparations without the interference of the erythrocytes. This technique is simple and convenient to prepare the chromosome of other fishes or other animals for FISH.     

改进的植物染色体制片方法

文章以外来物种肿柄菊和濒危物种三棱栎的根尖和茎尖为材料,改进植物染色体制片技术的结果表明:改进后的方法能明显提高植物染色体制片的质量和所需目的细胞的获得率。     

果蝇唾腺染色体的观察和制片方法的改进

几年来我们在教学中进行了多方面的实验探索,在果蝇唾腺制片方法上加以改进,采用加蔗糖液保色,树脂胶封边,制备一种半永久制片标本。操作过程如下: 1.取出唾腺,在其上加2滴(37℃)蒸馏水低渗处理10～20分钟,使唾腺细胞涨大有利于染色体的分散。 2.用滤纸吸去蒸馏水,加1滴1N HCl解离8～15分钟(时间随当时的温度而定),吸去解离液,用蒸馏水轻轻冲洗2～3次,将水吸净。     

植物染色体制片方法的改进

染色体是生物细胞核中最重要而稳定的成分，它具有特定的形态结构和一定的数目，具有自我复制能力，并积极参与细胞的代谢活动，能出现连续而有规律的变化，是决定物种繁衍的遗传物质的载体。人们一直在不断地探索和改进染色体的观察和鉴定方法。从早期的涂片法．到经典的常规压片法，直至20世纪60年代末建立起来的显带技术，以及90年代迅速发展的染色体原位杂交技术。作者长期从事植物染色体的研究，并给本科生和研究生讲授细胞遗传学理论和实验课，结合科研对染色体的制片技术进行了改进．供参考．[第一段]     

一种新的染色体制片永久封片法

细胞的有丝分裂和减数分裂制片是植物细胞学和细胞遗传学研究不可缺少的技术。对于一些重要的染色体制片进行永久封片,不仅为实验结果保存了必要的细胞学证据,而且也为特殊细胞学现象的重新观察和研究提供了可能性。传统染色体制片的封片剂主要有加拿大树脂(Canada balsam)、Euparal胶和DPX     

巴西橡胶树染色体制片方法的改良及FISH信号检测

本研究以巴西橡胶树热研7-33-97品种幼叶为材料,通过改良配制纤维素酶与果胶酶混合酶的溶剂、酶解时间等参数,对酶解去壁低渗方法进行了相应改良。结果表明,在37℃下,采用磷酸盐缓冲液(NaH2PO4,31.21 g/L; Na2HPO4, 71.64 g/L; Na Cl, 8 g/L; pH=5.5)作为溶剂,配制终浓度为5%纤维素酶、4%果胶酶的酶混合液时,酶解时间为1.0 h,大大缩短了酶解时间。用该法制备的中期标本,染色体形态分散、清晰、细胞膨大良好,荧光原位杂交信号检测后,信号正常。本研究可为橡胶树细胞遗传学、基因定位提供细胞学方法。     

介绍一种叶螨染色体的制片技术

<正> 叶螨总科(Tetranychoidea)包括了许多世界性分布的农业大害虫。研究叶螨的染色体,对于其分类、进化、性比和孤雌生殖等方面的研究均有十分重要的意义。叶螨个体小,染色体十分短小(一般1～4μm),给观察和研究带来了一定的难度。国外一般采用普通压片法制片、观察叶螨的染色体,但国内尚未见有关这方面的报道。作者以朱砂叶螨Tetranychuscinnabarinus(Boisduval)和二斑叶螨T.ur-ticae Koch为材料对叶螨染色体的制片技术进行了摸索,现简要报道如下。     

鱼类染色体组工程及其应用

鱼类具有产卵量大、体外受精的特点,是进行染色体组工程(Genome Engineering)研究的理想材料。运用染色体组工程技术可诱导鱼类的雌核发育(Gynogenesis)、雄核发育(Androgenesis)和多倍体(Polyploid)。雌核发育、雄核发育不仅可以用于建立近交系、生物学基础理论研究,而且还可以用于鱼类的单性养殖。多倍体在提高鱼类的生长率、成活率、抗病力以及种群控制等方面具有很大潜力。     

An Improved Method for Chromosome Preparations from Preimplantation Mammalian Embryos, Oocytes or Isolated Blastomeres

A method is described for making chromosome preparations from mammalian oocytes or preimplantation embryos, with or without the zona pellucida, or from isolated blastomeres. It is more robust and requires less skill and experience than previous techniques, yet chromosome structure is well preserved and very high quality preparations can be made. The method, which involves use of cold hypotonic solution and very cold fixative, reduces turbulence and allows even single blastomeres to be located and handled with relative ease, while the duration of hypotonic treatment becomes noncritical. The softening solution recommended contains no lactic acid and hence does not harm the chromosomes.     

Improved Methods for Permanent Chromosome Preparations: Cellophane Squashes and Citric Acid Dissociation

Normally, squash preparations from prestained acetic acid softened or HCl macerated tissues are made by pressing the tissue under a coverglass (e.g. Walker 1973). When permanent slides are wanted the coverglass has to be removed sooner or later, which is only possible by hardening the squashed tissue by freezing, for example in carbon dioxide snow, or by slow diffusion of fixatives into the space between the slide and the coverglass. These methods are either expensive or time-consuming, and upon removal of the coverglass, many cells and chromosomes either are lost or are poorly preserved.     

An Improved Fixing Solution for Methylene Blue Preparations

A method is described which combines the writer's hot celloidin technic¹ with a form of the clearing-before-cutting procedure. The method requires only 16–17 days and yields a block which may be cut in any microtome, the sections being as thin as those afforded by paraffin with comparable material. The advantages of celloidin over paraffin, listed in the writer's earlier paper, are retained in the present method which, altho consuming more time than the hot process, requires less skill and gives superior results.     

An In Vivo Giemsa Chromosome Banding Technique

An in vivo chromosome banding technique has been developed. Swiss albino mice were injected with the DNA alkylating agents ethyl methanesulfonate, methyl methanesulfonate, or methyl ethanesulfonate 12, 24, 48 or 72 hours prior to cell harvesting. After harvesting, the cells were fixed with 3:1 methanol-acetic acid and slides were prepared by air drying. The slides were stained 2¹/₂ minutes in 3% Giemsa in pH 6.8 Sorensen's buffer. All three alkylating agents induced chromosome bands similar to the Giemsa bands induced by other banding techniques which involve postfixation treatments.     

An Improved Method for Chromosome Counting in Maize

An improved method for counting chromosomes in maize (Zea mays L.) is presented. Application of cold treatment (5C, 24 hr), heat treatment (42 C, 5 min) and a second cold treatment (5C, 24 hr) to root tips before fixation increased the number of condensed and dispersed countable metaphase chromosome figures. Fixed root tips were prepared by the enzymatic maceration-air drying method and preparations were stained with acetic orcein. Under favorable conditions, one preparation with 50-100 countable chromosome figures could be obtained in diploid maize using this method. Conditions affecting the dispersion of the chromosomes are described. This technique is especially useful for determining the somatic chromosome number in triploid and tetraploid maize lines.     

淡水涡虫染色体的制备方法

以涡虫的再生组织为材料,用秋水仙素处理长有再生组织的涡虫片段,并通过改善各种实验条件,得到高清晰度的染色体图谱。结果表明,本方法简便易行,制成的染色体分散好,形态清晰,适用于对涡虫染色体的进一步研究。     

Chromosome Preparations from the Avian Allantoic Sac

An abundance of mitotic cells, a rapid and uniform response of cells to mitotic inhibition, ease in obtaining a monolayer of cells, clear and well-spread chromosomes make the allantois an ideal tissue for squash preparations. After a 45 min incubation of each embryo, still in the shell, with 0.02 ml of 0.05% Colcemid, 4-day avian embryos were treated in distilled water for 15 min or in 0.9% sodium citrate for 30-60 min and then fixed for at least 1 hr in 1:3 acetic-alcohol. Squash preparations were made after immersion of the allantois in 45% acetic acid for 5-10 mm. Phase contrast microscopy could be used, or permanent preparations made by freezing, air-drying and staining. Staining with Gram's iodine for 4 min followed by 1% crystal violet in 95% ethanol for 3 min is recommended. The allantois is well suited for use in biology laboratories to demonstrate avian chromosomes in different stages of mitosis.     

Simplified Culture and Chromosome Preparations of Primate Leukocytes

Plasma recovered from 1 ml of primate peripheral blood by centrifugation is planted in a medium consisting of 80% TC-199 and 20% fetal bovine serum, to which 0.125 ml of phytohaemagglutinin/5 ml is added. The pH is adjusted to 7 with 10% NaHCO₂. The mixture is incubated 68 hr, Colcemide to give 1 μg/ml is added, and incubation continued for 4 hr. Following centrifugal separation, the cells are given a hypotonic treatment with 0.75% sodium citrate for 15 min, then centrifuged again and fixed in 3:1 methanol-glacial acetic acid, 3 changes. Tiny drops of the cell suspension are placed on a slide, spread by blowing, and air dried. The preparations are stained with 15% Giemsa solution in methyl alcohol. The method has been successfully used in 256 specimens from 25 different species.