FISH参考

fish判读方法及标准
鱼类的判读方法及标准可以从多个方面来进行讨论。

首先，我们可以从外部特征、内部结构和生活习性等方面来进行判读。

从外部特征来看，鱼类的判读可以包括身体形状、鳞片特征、鳍的位置和形状、口的大小和位置等方面。

不同种类的鱼类在这些外部特征上会有明显的差异，通过对这些特征的观察和比对，可以初步判读出鱼类的种类。

内部结构也是判读鱼类的重要依据之一。

通过解剖鱼类，可以观察其骨骼结构、内脏器官的位置和形态等特征，这些都可以帮助确定鱼类的种属。

此外，鱼类的生活习性也是判读的重要依据之一。

不同种类的鱼类在生活习性上会有所不同，比如栖息环境、食性习惯、繁殖方式等。

通过对这些生活习性的了解和观察，也可以帮助判读鱼类的种类。

在实际判读中，还需要参考相关的鱼类分类学知识和专业的鱼类鉴定手册，以确保判读的准确性和科学性。

此外，对于商业和食
用鱼类，还需要参考相关的食品安全标准和法规，以确保所判读的
鱼类符合食品安全要求。

总的来说，鱼类的判读方法及标准是一个综合性的过程，需要
结合外部特征、内部结构、生活习性等多个方面的信息来进行判读，同时需要参考相关的鱼类分类学知识和食品安全标准，以确保判读
的准确性和科学性。

HER2基因荧光原位杂交(FISH)检测是乳腺癌诊断和治疗中的重要方法。

HER2基因在某些乳腺癌患者中会发生扩增，导致肿瘤的侵袭性增强。

FISH检测可以检测HER2基因是否存在扩增，从而指导患者的治疗方案。

HER2基因FISH检测的判读标准通常基于以下几个方面：
1. HER2基因信号数:在FISH图像中，HER2基因通常呈现为成簇的红色信号和一个绿色的对照信号。

根据判读标准，红色信号的数量应该与绿色信号的数量相等或更多。

如果红色信号的数量少于绿色信号的数量，则可能表示HER2基因没有扩增或扩增程度较低。

2. HER2/CEP17比值:FISH图像中，HER2基因和CEP17(17号染色体的特定区域)通常会同时呈现为红色和绿色信号。

通过计算HER2信号和CEP17信号的比值，可以估算出肿瘤细胞中HER2基因的拷贝数。

如果比值大于等于2.0，则通常表示HER2基因存在扩增；如果比值小于2.0，则通常表示HER2基因没有扩增或扩增程度较低。

3. 平均HER2信号强度:除了计算HER2/CEP17比值之外，还可以单独评估HER2基因的平均信号强度。

如果平均信号强度明显高于背景噪声，则可能表示HER2基因存在扩增；如果平均信号强度与背景噪声相当或较低，则可能表示HER2基因没有扩增或扩增程度较低。

需要注意的是，FISH检测结果的判读应由经验丰富的实验室技术人员进行，并结合其他临床和病理学信息来综合评估患者的病情。

此外，不同的实验室和检测方法可能会有不同的判读标准，因此在比较不同实验室的结果时需要谨慎。

★英文写作翻译频道为大家整理的我最喜欢的动物英语作文带翻译鱼，供大家
参考!
最近我养了一条麦穗鱼，特别可爱，呵呵。

所以在这里为大家提供篇关于鱼的英语作文吧。

希望您也能跟我一起分享其中的快乐-作文地带。

关于我最喜欢的动物是鱼的小学生英语作文
My favourite animal is fish.The fish are very cute.
我最喜欢的动物是鱼，鱼特别可爱(乖巧)。

There are many kinds of fish,such as,Goldfish,the beautiful fish,many people like to feed them.Tropical fish,they live in the sea.Whale,They are the largest animal in the world.
鱼的种类有很多种，比如金鱼、观赏鱼，很多人都喜欢饲养鱼。

热带鱼生活在大海中，他们是世界上的动物(这个作文地带不大认同)，作者估计是要说，鱼是世界上种类最多的动物。

They fish sleep without close eyes.Some fish sleep during the day , and some fish sleep at night.Generally sleep through out the night.
鱼睡觉从来不闭眼的。

一些鱼白天睡觉，一些鱼晚上睡觉。

一般来说，鱼都是晚上睡眠的。

fish判读方法及标准全文共四篇示例，供读者参考第一篇示例：鱼类是一种常见的食物，不仅味道鲜美，而且富含蛋白质和多种维生素，对人体健康有益。

鱼类易腐烂，容易受到污染，所以在购买和食用鱼类时需要注意判断其新鲜程度。

下面将介绍一些关于鱼类新鲜度判断方法及标准。

判断鱼类的外观。

新鲜的鱼类应该外表光滑，有光泽，鳞片密集，没有褪色或变色的现象。

鱼眼应该凸出且透明，没有浑浊的情况。

鱼鳍应该完整，没有断裂或变形。

鱼腹部应该有一层薄薄的透明膜，如果这个膜变浑浊或发黄，则表示鱼不新鲜。

鱼类应该有一股清新的海味，如果有异味或腥味，则表示鱼不新鲜。

判断鱼类的触感。

新鲜的鱼应该肉质有弹性，按压时有一定的回弹感。

如果鱼肉感觉松软或触感不良，则表示鱼不新鲜。

可以用手指轻轻按压鱼肉，看是否有明显的油脂流出。

如果没有油脂或者油脂呈现混浊状态，则表示鱼类不新鲜。

判断鱼类的气味。

新鲜的鱼应该有清新的海味，没有异味或腥味。

可以用鼻子贴近鱼体闻一下，如果有刺鼻的气味或异味，则表示鱼类不新鲜。

可以用手摸一下鱼鳃，看鱼鳃是否有强烈的氨味，如果有，则表示鱼不新鲜。

判断鱼肉的色泽。

新鲜的鱼应该具有鲜艳的颜色，比如鲜红色的金枪鱼肉或鲜粉色的鲑鱼肉。

如果鱼肉颜色很暗淡或发黑，则表示鱼类不新鲜。

还可以用手指轻轻压一下鱼肉，看是否会留下明显的印痕。

如果留下较深的印痕或者鱼皮因压力而裂开，则表示鱼类不新鲜。

判断鱼类的新鲜度应该综合考虑外观、触感、气味和色泽等方面。

如果鱼类在这些方面表现良好，则证明其新鲜度较高，可以放心购买和食用。

相反，如果在以上方面存在异常情况，则应慎重考虑是否购买这种鱼类。

只有通过仔细观察和判断，我们才能选择到新鲜的鱼类，为自己和家人健康饮食提供保障。

第二篇示例：鱼类是一种重要的动物资源，除了作为食用品外，还具有重要的经济和生态价值。

对于鱼类进行检验和判读是至关重要的。

在实际操作中，鱼类的判读方法及标准是非常重要的，下面我们将详细介绍。

Video ArticleFluorescent in situ Hybridization on Mitotic Chromosomes of MosquitoesVladimir A. Timoshevskiy1, Atashi Sharma1, Igor V. Sharakhov1, Maria V. Sharakhova11Department of Entomology, Virginia TechCorrespondence to: Maria V. Sharakhova at msharakh@URL: /video/4215DOI: doi:10.3791/4215Keywords: Immunology, Issue 67, Genetics, Molecular Biology, Entomology, Infectious Disease, imaginal discs, mitotic chromosomes, genome mapping, FISH, fluorescent in situ hybridization, mosquitoes, Anopheles, Aedes, CulexDate Published: 9/17/2012Citation: Timoshevskiy, V.A., Sharma, A., Sharakhov, I.V., Sharakhova, M.V. Fluorescent in situ Hybridization on Mitotic Chromosomes of Mosquitoes. J. Vis. Exp. (67), e4215, doi:10.3791/4215 (2012).AbstractFluorescent in situ hybridization (FISH) is a technique routinely used by many laboratories to determine the chromosomal position of DNA and RNA probes.One important application of this method is the development of high-quality physical maps useful for improving the genome assemblies for various organisms.The natural banding pattern of polytene and mitotic chromosomes provides guidance for the precise ordering and orientation of the genomic supercontigs.Among the three mosquito genera, namely Anopheles, Aedes, and Culex, a well-established chromosome-based mapping technique has been developed only for Anopheles, whose members possess readable polytene chromosomes 1. As a result of genome mapping efforts, 88% of the An. gambiae genome has been placed to precise chromosome positions 2,3 . Two other mosquito genera, Aedes and Culex, have poorly polytenized chromosomes because of significant overrepresentation of transposable elements in their genomes 4, 5, 6. Only 31 and 9% of the genomic supercontings have been assigned without order or orientation to chromosomes of Ae. aegypti 7 and Cx.quinquefasciatus 8, respectively. Mitotic chromosome preparation for these two species had previously been limited to brain ganglia and cell lines. However, chromosome slides prepared from the brain ganglia of mosquitoes usually contain low numbers of metaphase plates 9. Also, although a FISH technique has been developed for mitotic chromosomes from a cell line of Ae. aegypti 10, the accumulation of multiple chromosomal rearrangements in cell line chromosomes 11 makes them useless for genome mapping. Here we describe a simple, robust technique for obtaining high-quality mitotic chromosome preparations from imaginal discs (IDs) of 4th instar larvae which can be used for all three genera of mosquitoes. A standard FISH protocol 12 is optimized for using BAC clones of genomic DNA as a probe on mitotic chromosomes of Ae. aegypti and Cx.quinquefasciatus, and for utilizing an intergenic spacer (IGS) region of ribosomal DNA (rDNA) as a probe on An. gambiae chromosomes. In addition to physical mapping, the developed technique can be applied to population cytogenetics and chromosome taxonomy/systematics of mosquitoes and other insect groups.Video LinkThe video component of this article can be found at /video/4215/Protocol1. Chromosome PreparationMosquito larvae were reared using a standard protocol described in Methods in Anopheles Research available at the website of the Malaria Research and Reference Reagent Resource Center(MR4) 13. The temperatures of mosquito rearing were modified to provide the highest number of chromosomes in imaginal discs and lowest mortality of the larvae. The stages of mosquito larvae development were determined based on the sizes of their head capsules 13.1.Hatch mosquito eggs at 28 °C, and after 2-3 days, transfer 2nd or 3rd instar larvae to 16 °C for Ae. aegypti and Cx. quinquefasciatus and to 22 °C for An.gambiae.2.Place 4th instar larvae on ice for several minutes for immobilization.3.Transfer larva to a slide with a drop of cold hypotonic solution (0.5% sodium citrate or 0.075 M potassium chloride), and place it under the stereomicroscope.4.Select larva with oval IDs (Figure 1B) for further dissection.5.Decapitate larva, and cut the cuticle from the ventral side of the larval thorax using dissecting scissors (Figure 2A). Make additional cut in second or thirdabdominal segment to dissect the gut from the larva. The directions of the cuts are shown by arrows.6.Open the cuticle, and remove the gut and fat body from the larva. Remove the hypotonic solution from the slide using filter paper, and add a fresh drop ofhypotonic solution directly to the IDs (Figure 2B). Keep larva in hypotonic solution for 10 min at RT.7.Remove hypotonic solution using filter paper, and apply Carnoy's solution (ethanol/acetic acid in 3:1 ratio). After adding fixative solution, IDs immediatelyturn white and become easily visible under the microscope (Figure 2C).ing dissecting needles, remove IDs from the larva (Figure 2D), and transfer them to a drop of 50% propionic acid. Remove any other tissues, such as thegut and fat body, from the slide. Cover IDs with an unsiliconized 22x22 cover slip, and keep for 10 min at RT.9.Cover the slide with filter paper, and squash the tissue by tapping the eraser of a pencil on the perimeter of the cover slip.10.Briefly analyze the quality of the slide using the phase-contrast microscope at 100x or 200x magnification (Figure 3). Preparations with >50 chromosomespreads can be considered suitable for FISH.11.Dip and hold the slide in liquid nitrogen until it stops bubbling. Remove the cover slip from the slide using a razor blade, and transfer the slide immediatelyto a container of 70% ethanol chilled at -20 °C. Store at 4 °C for at least 1 hr for the best dehydration result (if necessary, slides can be stored at this step from several minutes to several days).12.Dehydrate slides in a series of ethanol (70%, 80%, 100%) at 4 °C for 5 min each, and air dry at RT.13.Store dry slides at -20 °C before utilizing them for FISH.2. Extraction of Repetitive DNA FractionsPerforming FISH of the BAC clone DNA probe on chromosomes from Ae. aegypti and Cx. quinquefasciatus requires using unlabeled repetitive DNA fractions to block unspecific hybridization of the DNA repeats to the chromosomes. The reassociation of single-strand DNA fragmented into pieces of several hundred bp follows a C0t curve where C0 is the initial concentration of single-stranded DNA and t is the reannealing time. DNA fractions with C0t values equal to 10-4-10-1 or 10°-102 are considered as highly and moderately repetitive, respectively.1.Extract 400-500 µg of the genomic DNA from entire adult mosquito using Qiagen Blood and Cell Culture Maxikit, and prepare 100-1,000 ng/µl DNAsolution in 1.2x SSC.2.Denature DNA by placing a safe-lock tube with genomic DNA into a heating block prewarmed to 120 °C for 2 min. High temperature helps to range DNAinto 200-500 bp fragments.3.Depending on the DNA concentration, reassociate DNA by placing the tube at 60 °C for 15-150 min to obtain C0t DNA fractions up to C0t3 (Table 1).4.Place the tube with DNA on ice for 2 min.5.Transfer the DNA to 42 °C, add preheated 10x S1 nuclease buffer and S1 nuclease to a final concentration of 100 U per 1 mg of DNA, and incubate for 1hr.6.Precipitate DNA by adding 0.1 volume of 3 M sodium acetate and 1 volume of isopropanol at RT.7.Centrifuge at 14,000 rpm for 20 min at 4 °C.8.Wash DNA in 70% ethanol, and centrifuge again at 14,000 rpm for 10 min at 4 °C.9.Air-dry and dissolve DNA pellet in TE buffer.10.Measure the DNA concentration, and visualize by gel electrophoresis. Usually the final quantity of repetitive DNA fractions represents 35-50% of theoriginal DNA amount.3. DNA Probe LabelingTwo different protocols were used for the labeling BAC clone DNA probe and IGS rDNA probe.3.1 BAC clone labeling using nick-translation1.Extract BAC clone DNA from the BAC library using Qiagen Large Construct Kit.2.Prepare reaction mixture for nick-translation labeling on ice with final volume of 50 µl: 1 µg isolated BAC clone DNA, 0.05 mM each of unlabeled dATP,dCTP, and dGTP and 0.015 mM of dTTP; 1 µl of Cy3-dUTP (or another fluorochrome); 0.05 mg/ml of BSA, 5 µl of 10x nick-translation buffer, 20 U of DNA-polymerase I, and 0.0012 U of DNase.3.Incubate at 15 °C for 2.5 hr.4.Stop reaction by adding 1 µl of 0.5 M EDTA.5.Store probe at -20 °C in a dark place.3.2 IGS rDNA labeling using PCR1.Prepare reaction mixture on ice with final volume of 50 µl: 200 ng of genomic DNA; 0.05 mM each of unlabeled dATP, dCTP, and dGTP; 0.015 mM ofdTTP; 1 µl of Cy3-dUTP (or another fluorochrome); 5 µl of 10x PCR-buffer; 50 pmol of forward; UN (GTGTGCCCCTTCCTCGATGT) and reverse; GA (CTGGTTTGGTCGGCACGTTT) primers for IGS amplification; and 10 U of Taq DNA polymerase 14.2.Perform PCR reaction using standard PCR parameters for IGS amplification: 95 °C /5 min x 1 cycle; (95 °C /30 sec, 50 °C /30 sec, 72 °C /30 sec) x 30cycles; 72 °C /5 min x 1 cycle; and 4 °C hold 14.3.Store probe at -20 °C in a dark place.4. Fluorescent in situ HybridizationThis FISH protocol includes two variations: the first for using BAC clone DNA as a probe on mitotic chromosomes of Ae. aegypti and Cx. quinquefasciatus and the second for using IGS rDNA on mitotic chromosomes of An. gambiae. If using BAC clone DNA probes, skip RNase treatment steps 4.3, 4.4, and simultaneous slide/probe denaturation step 4.19. If using IGS rDNA probe, prepare hybridization mixture without C0t DNA fractions, and skip separate slide/ probe denaturing steps 4.10, 4.11, 4.16, and 4.17.1.Incubate slides in 2x SSC for 30 min at 37 °C.2.Dehydrate slides in series of 70%, 80%, and 100% ethanol for 5 min each at RT, and air dry. If performing FISH with BAC clone DNA, proceed directly tostep 4.5.3.Incubate chromosome preparation in 0.1 mg/ml RNase solution under parafilm for 30 min at 37 °C.4.Wash twice in 2x SSC for 5 min each at 37 °C.5.Put slides in a jar with 0.01% pepsin and 0.037% HCl solution, and incubate for 5 min at 37 °C.6.Wash slides in 1x PBS for 5 min at RT.7.Fix chromosome preparation in a jar with 1% formalin in 1x PBS prepared from 10% neutral-buffered formalin for 10 min at RT.8.Wash slides in 1x PBS for 5 min at RT.9.Dehydrate slides in series of 70%, 80%, and 100% ethanol for 5 min each at RT, and air dry preparations at 37 °C. If performing FISH with IGS, proceeddirectly to step 4.1210.Denature slides in a jar with prewarmed 70% formamide for 2 min at 72 °C.11.Dehydrate slides in series of cold (-20 °C) 70%, 80%, and 100% ethanol for 5 min each, and air dry at 37 °C.12.Prepare hybridization mixture: 5 µl of labeled probe DNA from step 3, 10 µl of C0t DNA from step 2 with final concentration of 0.5 ng/µl, and 5 µl of 1 µg/µl sonicated salmon sperm DNA. For FISH with IGS rDNA, prepare hybridization mixture without C0t DNA fractions.13.Precipitate DNA by adding 0.1 volume of 3 M sodium acetate and 2 volumes of ethanol. Keep at -20 °C for 1-3 hr.14.Centrifuge at 14,000 rpm at 4 °C for 20 min, remove the ethanol, and air dry the pellet at RT.15.Thoroughly dissolve the pellet in 10 µl of hybridization buffer: 50% formamide, 20% dextran sulfate, 2x SSC. If performing FISH with IGS, proceeddirectly to step 4.1816.Denature hybridization mixture for 7 min at 97 °C, and immediately put on ice for 1 min.17.Prehybridize mixture at 37 °C for 30 min to prevent unspecific hybridization of repetitive DNA to the chromosomes.18.Place 10 µl of the hybridization mixture on the slide, and cover with a 22x22 cover slip. Prevent bubble formation - air bubbles should be removed withgentle pressure to the coverslip. If performing FISH with BAC clone DNA, proceed directly to step 4.2019.Denature the probe and chromosome DNA simultaneously using a heating block at 75 °C for 5 min.20.Glue cover slip around the perimeter using rubber cement.21.Perform overnight hybridization in a humid chamber at 37 °C.22.Remove rubber cement and coverslip from the slide.23.Wash slide 2 min in prewarmed Solution 1 (0.4x SSC, 0.3% Nonidet-P40) at 73 °C.24.Wash slides in Solution 2 (2x SSC, 0.1% Nonidet-P40) for 5 min at RT.25.Counterstain slide using 0.001 mM YOYO-1 in 1x PBS for 10 min in humid chamber at RT.26.Mount in a small amount of Prolong Gold antifade reagent with a cover slip.27.Analyze preparations under a fluorescent microscope using appropriate filter sets at 1,000x magnification (Figure 4).5. Representative ResultsInsect IDs are located in each segment of the larva. Depending on the position, they transform into different tissues at the adult stage of the insect. The IDs, which are used for the chromosome preparation in this protocol, develop into legs at the adult stage of the mosquito. These IDs are located at the ventral side of the larval thorax and are clearly visible through the cuticle under the microscope (Figure 1). At the early 4th instar larval stage, IDs have a round shape (Figure 1A). The largest numbers of mitosis, ~175 in one ID 9, are accumulated at a later "oval shaped" stage (Figure 1B), which must be considered the optimal stage for slide preparation. At this time, the intermediate ID splits into two: one transforms into a leg and another one transforms into a wing. We prefer using the large leg IDs at the "oval-shaped" stage for the chromosome slide preparation. Figure 1C represents IDs at the latest stage of 4th instar larva development. At this stage, the IDs are already developed into legs and wings, and contain a significant amount of differentiated tissues and a low number of mitosis. This stage of ID development should be avoided for chromosome slide preparation. We also recommend rearing mosquito larvae at low temperatures: 16 °C for Aedes and Culex and 22 °C for Anopheles. This helps to increase the amount of mitosis in IDs 9.Figure 2 illustrates ID dissection from the thorax of 4th instar larva. Because the cuticle of a live insect is hard to dissect, we recommend using dissecting scissors instead of the needles commonly used for larva preparation. The most crucial procedure for obtaining high-quality chromosome preparation is the hypotonic solution treatment. For best results, we remove the gut and fat body from the larval thorax before this treatment. Swelling of the ID cells during this procedure helps to spread chromosomes on a slide (Figure 3A). The appropriate quality of the hypotonic solution treatment can be easily recognized by the round shape of cells in the preparations (Figure 3A, B). Cells with an oval shape indicate insufficient hypotonic solution treatment (Figure 3C). To be selected for FISH, chromosome preparation should contain at least 50 high-quality chromosome spreads. Normally, ~90% of the slides prepared using this protocol have sufficient quality for FISH 9.We present two slightly different FISH protocols: an advanced protocol for FISH using genomic BAC clone DNA probe on mitotic chromosomes of Aedes and Culex and a simple FISH protocol for IGS rDNA probe on mitotic chromosomes of Anopheles. The genomes of Aedes and Culex are highly repetitive because of the overrepresentation of transposable elements 7,8. Thus, performing FISH, which utilizes genomic BAC clone DNA as a probe, requires adding unlabeled repetitive DNA fractions to the probe to block unspecific hybridization of the DNA repeats to chromosomes. For the extraction of the repetitive DNA fractions, genomic DNA is denatured at 120 °C for 2 min. Boiling DNA at a high temperature also helps to obtain DNA in fragments of 200-500 bp. DNA is allowed to reassociate after this treatment. The highly repetitive DNA fragments tend to find their mate for reassociation faster than DNA with unique sequences does. As a result, the reassociation of DNA follows a C0x t curve where C0 is the initial concentration of single-stranded DNA, and t is the reannealing time.DNA fractions with C0t values equal to 10-4--10-1 or 100-102 are considered highly and moderately repetitive, respectively. The time of the reassociation for different C0t DNA fractions can be calculated using the formula t= C0t X × 4.98/C0 , where t - time of incubation, C0t X - C0t fraction (C0t1=1, C0t2=2, etc.) and C0 - initial DNA concentration in µg/µl15 (Table 1). After reassociation, the single-stranded DNA is digested using S1 nuclease. We prefer using all C0t DNA fractions up to C0t3 together instead of the commonly used C0t1 DNA fraction. These C0t fractions include some of the moderately repetitive DNA sequences and together usually represent 35-50% of the original amount of the genomic DNA in Ae. aegypti. The correct proportion between labeled DNA probe and unlabeled C0t DNA fraction depends on the repetitive DNA component in each particular BAC clone. On average, we use 1:20 probe to C0t DNA fraction proportion for obtaining an acceptable signals/background ratio of the FISH result. Prehybridization of the DNA probe with C0t DNA fractions in a tube for 30 min before the actual hybridization on the slide also helps to reduce background. Labeling, hybridization itself, and washing in this protocol are performed using standard conditions 12.The FISH results of two differently labeled BAC clone DNA probes on mitotic chromosomes of Ae. aegypti and Cx.quinquefasciatus are shown in Figures 4A and B, respectively. The BAC clone DNA probes produce strong signals in a single position on the chromosomes. Chromosomes shown in Figure 1 are counterstained with YOYO-1 iodide. This dye produces the best banding patterns on Ae. aegypti chromosomes 9. Alternatively, other fluorescent dyes, such as DAPI or propidium iodide, can be utilized for the chromosome counterstaining. For suppressing photobleaching of the slides, we use Prolong Gold antifade mounting medium. This reagent has good signal preservation abilities and also can be easily removed from the slide by rinsing in 1x PBS if it is necessary to use the same slide for several hybridizations.A simple version of the FISH protocol is designed for hybridization of IGS rDNA probe on mitotic chromosomes of Anopheles. Ribosomal genes in Anophelesare represented as a polymorphic cluster of genes located on sex chromosomes 16. A DNA probe in this protocol is labeled using standard PCR reaction by adding fluorescently labeled Cy3 or Cy5 dNTPs. Because blocking unspecific hybridization of repetitive DNA in euchromatin is not needed, all steps related to using C0t DNA fractions are omitted. Instead, chromosome preparations are pretreated with RNase for preventing hybridization of the IGS rDNA probe to thenucleolus. Chromosomes and the DNA probe are denatured simultaneously by heating the slide together with a probe in a hybridization system at 75 °C for 5min. Hybridization and washing in this protocol are also performed using standard conditions for FISH 12. The result of FISH is demonstrated in Figure 4C: the polymorphism of the IGS rDNA hybridization between two X chromosomes is clearly visible.DNA concentration µg/µl Reannealing time,min0.11000.3330.5200.7140.911C 0t 21100.11500.3500.5300.7210.917C 0t 3115Table 1. DNA concentration and reannealing times for preparation of C 0t2 and C 0t3 fractions.Figure 1. Stages of the ID development in 4th instar larva: A) an early "round shape" stage; B) an intermediate "oval shape" stage - optimal for thechromosome preparation; C) a late stage - inappropriate for chromosome preparations. The positions of IDs are indicated by arrows on the ventral side of the larval thorax.Figure 2. Steps of ID dissection: A) decapitated larva (the direction of cuts are indicated by arrows); B) larvae with dissected gut under hypotonic solution treatment (IDs swell and become almost invisible); C) larva after Carnoy's solution application (IDs become white and clearly visible); D) dissected IDs in Carnoy's solution. Positions of IDs in larva are indicated by asterisks.Figure 3. Different qualities of the chromosome spreads: A) a perfect chromosome spread - round shape of the cells demonstrates sufficient treatment of the IDs in hypotonic solution; B) a perfect hypotonic treatment - chromosomes are slightly undersquashed; C) a poor chromosome spread - the result of insufficient hypotonic treatment is indicated by oval shape of the cells.Figure 4. Examples of FISH with BAC clones (A, B) and IGS rDNA (C) in the chromosomes of Ae. aegypti (A), Cx.quinquefasciatus (B), and An. gambiae(C). 1, 2 and 3 - are numbers of chromosomes; X - female sex chromosome in An. gambiae.DiscussionNonfluorescent in situ hybridization on mitotic chromosomes of mosquitoes was performed for the first time in 1990 by A. Kumar and K. Rai 17. In that study, 18S and 28S ribosomal DNA genes, cloned together in one plasmid, were placed to the chromosomes of 20 species of mosquitoes. The DNA probe was radioactively labeled and hybridized to the chromosomes from brain ganglia. Among three mosquito genera, a FISH technique has been developed only for mitotic chromosomes from the cell line of Ae. aegypti10,18,19 and has never been performed on mitotic chromosomes from live mosquitoes. Recently, we developed a simple, robust technique for obtaining high-quality chromosome preparations from IDs of 4th instar larvae 9. This method allows a high number of chromosomes to be obtained in one slide and can be universally used for all species of mosquitoes. The necessity of using only larval, not pupal or adult stages of mosquitoes, for slide preparation is probably the only limitation of the method. The standard FISH method 12 was optimized for using genomic BAC clone and IGS rDNA as probes for the mitotic chromosomes of Aedes, Culex, and Anopheles.In addition to these specific applications, the FISH protocols described here can also be used for other purposes. The advanced FISH protocol, which utilizes C0t DNA fractions for blocking unspecific hybridization, can also be applied for the hybridization of BAC clones or any other large DNA fragments in heterochromatic regions of Anopheles. Heterochromatic regions are enriched with transposable elements and other repeats, and probes from these regions normally produce strong background on the chromosomes3. Using unlabeled C0t DNA fractions will help to reduce unspecific hybridization of the probe to the chromosomes. The simple version of the FISH protocol can be used for any rDNA or repetitive DNA probes on mitotic chromosomes of mosquitoes and other insects. In addition, it also can be applied for the hybridization of BAC clone DNA in species with low repetitive DNA content in euchromatic regions such as Anopheles or Drosophila. The protocol proposed here will help to obtain highly-finished chromosome-based genome assemblies for mosquitoes and can be broadly used for various cytogenetic applications in other groups of insects.DisclosuresNo conflicts of interest declared.AcknowledgementsWe thank Sergei Demin and Tatyana Karamysheva for their help with chromosome preparation and FISH on Anopheles. We also thank David Severson for providing us Aedes and Culex genomic DNA BAC clones and Melissa Wade for editing the text. This work was supported by two grants from the National Institutes of Health: 1R21 AI88035-01 to Maria V. Sharakhova and 1R21 AI094289-01 to Igor V. Sharakhov.References1.Sharakhov, I.V. & Sharakhova, M.V. In: Chromosome Mapping Research Developments., Verrity, J.F. & Abbington, L.E., eds., Nova Science Publishers,Inc., (2008).2.Holt, R.A., et al. The genome sequence of the malaria mosquito Anopheles gambiae. Science.298, 129-149, doi:10.1126/science.1076181 (2002).3.Sharakhova, M.V., et al. Update of the Anopheles gambiae PEST genome assembly. Genome biology.8, R5, doi:10.1186/gb-2007-8-1-r5 (2007).4.Campos, J., Andrade, C.F., & Recco-Pimentel, S.M. A technique for preparing polytene chromosomes from Aedes aegypti (Diptera, Culicinae). Memoriasdo Instituto Oswaldo Cruz.98, 387-390 (2003).5.Campos, J., Andrade, C.F., & Recco-Pimentel, S.M. Malpighian tubule polytene chromosomes of Culex quinquefasciatus (Diptera, Culicinae). Memoriasdo Instituto Oswaldo Cruz.98, 383-386 (2003).6.McAbee, R.D., Christiansen, J.A., & Cornel, A.J. A detailed larval salivary gland polytene chromosome photomap for Culex quinquefasciatus (Diptera:Culicidae) from Johannesburg, South Africa. J. Med. Entomol.44, 229-237 (2007).7.Nene, V., et al. Genome sequence of Aedes aegypti, a major arbovirus vector. Science.316, 1718-1723 (2007).8.Arensburger, P., et al. Sequencing of Culex quinquefasciatus establishes a platform for mosquito comparative genomics. Science.330, 86-88, doi:10.1126/science.1191864 (2010).9.Sharakhova, M.V., et al. Imaginal discs--a new source of chromosomes for genome mapping of the yellow fever mosquito Aedes aegypti. PLoS neglectedtropical diseases.5, e1335, doi:10.1371/journal.pntd.0001335 (2011).10.Brown, S.E., et al. Toward a physical map of Aedes aegypti. Insect Mol. Biol.4, 161-167 (1995).11.Steiniger, G.E. & Mukherjee, A.B. Insect chromosome banding: technique for G- and Q-banding pattern in the mosquito Aedes albopictus. Can. J. Genet.Cytol.17, 241-244 (1975).12.Garimberti, E. & Tosi, S. In: Fluorescence in situ hybridization (FISH)., Bridger, J.M. & Volpi, E.V., eds., Springer Science and Business Media, (2010).13.Methods in Anopheles Research [Internet]. Atlanta (GA): The Malaria Research and Reference Reagent Resource Center. Available from: http:///Portals/3/Pdfs/ProtocolBook/MethodsAnophelesResearchV4c.pdf (2007).14.Scott, J.A., Brogdon, W.G., & Collins, F.H. Identification of single specimens of the Anopheles gambiae complex by the polymerase chain reaction. TheAmerican journal of tropical medicine and hygiene.49, 520-529 (1993).15.Trifonov, V.A., Vorobyeva, N N., & Rens, W. In: Fluorescence in situ hybridization (FISH)., Leiehr, T., ed., Spriger-Verlag, (2009).16.Collins, F.H., et al. A ribosomal RNA gene probe differentiates member species of the Anopheles gambiae complex. The American journal of tropicalmedicine and hygiene.37, 37-41 (1987).17.Kumar, A. & Rai, K.S. Chromosomal localization and copy number of 18S+28S ribosomal RNA genes in evolutionary diverse mosquitoes (Diptera,Culicidae). Hereditas.113, 277-289 (1990).18.Brown, S.E. & Knudson, D.L. FISH landmarks for Aedes aegypti chromosomes. Insect Mol. Biol.6, 197-202 (1997).19.Brown, S.E., Severson, D.W., Smith, L.A., & Knudson, D.L. Integration of the Aedes aegypti mosquito genetic linkage and physical maps. Genetics.157,1299-1305 (2001).。

· fish· v. [fɪʃ] ( fishes; fished; fishing )·· 双解释义· vt. & vi. 1.捕鱼; 钓鱼try to catch fish in a piece of water· vt. & vi. 2.摸出,掏出bring up· 基本要点•1.fish作动词的基本含义是指用工具“钓鱼”“捕鱼”“从…中捕捞”,引申还可表示“摸出(某种东西)”“掏出(某种东西)”。

2.fish可用作及物动词,也可用作不及物动词。

用作及物动词时,可接河、湖、溪、池等名词作简单宾语。

可用于被动结构。

•· 词汇搭配••fish contentedly 高兴地钓鱼•fish intently 专心地钓鱼•fish placidly 静静地钓鱼•fish around 翻来翻去•fish out 从水中把…捞出•fish out from 从…里捞出•fish up 从…里拖出••fish for a living 以捕鱼为主•fish for information 探听消息•fish in the air 缘木求鱼•fish in the sea 在海里捕鱼•fish on a stream 在小溪捕鱼•fish out of 从…里捞出•fish through the ice 破冰捕鱼· 常用短语•fish for(v.+prep.)▲fish for sth1.捕鱼 catch fishIt is no use fishing for trout in this river, there are none left.这条河里捕不到鲑鱼了,连一条也没有了。

2.摸索着找 search for by touchHe fished for a coin in his pocket.他在口袋里摸着找一枚硬币。

钓鱼英语作文范文参考示例Fishing is a popular outdoor activity that I recently had the opportunity to try. It was a memorable experience that allowed me to reconnect with nature and learn new skills.Early one morning, armed with my fishing rod, bait, and a sense of excitement, I set out to a nearby lake. The calm water reflected the vibrant colors of the surrounding trees, creating a tranquil atmosphere. With each step, I could feel the anticipation building inside me.Setting up my equipment, I carefully attached the bait to the hook. As I cast my line into the water, I was mesmerized by the rhythmic movement of the floating lure. It danced along the surface, imitating a wounded prey and luring fish to bite.With patience being the key, I relaxed and enjoyed the soothing sounds of nature. The chirping of birds and thegentle rustling of leaves created a symphony that reminded me of the beauty surrounding me. Hours passed by, and although I had not yet caught anything, I remained determined.Suddenly, my fishing rod twitched, and I felt a strong tug on the line. This was it – the moment I had been waiting for! Adrenaline surged through my veins as I began reeling in the fish. With skillful guidance, I gradually brought it closer to the shore.As the fish emerged from the water, glistening in the sunlight, a sense of accomplishment washed over me. It was a beautiful specimen, its scales shimmering with vibrant hues. Carefully, I removed the hook and marveled at its sleek form before releasing it back into the water, allowing it to swim away freely.Reinvigorated by my first catch, my enthusiasm grew, and I continued casting my line. Throughout the day, I caught several more fish, each encounter providing a surge ofexcitement and a deep appreciation for the wonders of nature. Fishing allowed me to disconnect from the noise and distractions of everyday life, providing a meditative escape.As dusk settled in, I decided to pack up and head home. The sun began to dip beneath the horizon, casting a golden glow over the landscape. With tired muscles and a content heart, I left the lake, already eager for my next fishing adventure.Fishing had not only allowed me to engage in a new hobby but had also provided a much-needed break from the demands of daily life. The experience reminded me of the importance of slowing down, appreciating nature, and finding joy in the simplest of activities. Fishing had become a cherished pastime, offering a peaceful sanctuary amidst the chaos of the modern world.。

fish1检查报告解读对于fish1检查报告的解读，我将从多个角度给出全面的回答。

首先，fish1（Fluorescence In Situ Hybridization）是一种用于检测染色体异常的分子遗传学技术。

它通过使用荧光探针与特定染色体区域的DNA序列结合，可以直接在细胞核中观察到染色体的结构和数量变化。

在解读fish1检查报告时，需要注意以下几个方面：1. 检测目标，检查报告中应该明确指出所检测的染色体区域或基因。

这可以帮助确定是否存在染色体异常或基因突变。

2. 结果解释，报告中可能会给出阳性或阴性的结果。

阳性结果表示存在染色体异常或基因突变，而阴性结果则表示未检测到异常。

需要注意的是，阴性结果并不代表绝对正常，因为染色体异常可能存在于未检测到的区域。

3. 异常类型，如果报告显示阳性结果，应该详细说明检测到的染色体异常类型。

这可能涉及到染色体缺失、重复、重排或染色体数目异常等。

4. 异常程度，报告中可能会提供染色体异常的程度。

例如，染色体缺失的大小、重复的次数、重排的范围等。

这有助于评估异常对个体健康的影响。

5. 临床意义，解读报告时需要结合临床情况来评估染色体异常的临床意义。

染色体异常可能与某些遗传疾病、肿瘤等相关联，但并非所有异常都会导致疾病。

此外，解读fish1检查报告还需要考虑以下几个因素：1. 技术准确性，fish1技术虽然高度敏感和特异，但仍可能存在假阳性或假阴性结果。

因此，报告结果应该与其他检查方法进行比较和确认。

2. 个体差异，不同个体之间染色体结构和数量可能存在差异。

因此，需要将检测结果与正常参考范围进行比较，以确定是否存在异常。

3. 进一步检查，如果fish1检查报告显示染色体异常，通常需要进一步的检查来确认和评估异常的具体情况。

例如，核型分析、基因突变检测等。

总体而言，解读fish1检查报告需要综合考虑检测目标、结果解释、异常类型和程度、临床意义等因素。

同时，需要注意技术准确性和个体差异，并在必要时进行进一步的检查。

乳腺癌HER2基因FISH检测判读标准乳腺癌是人类最常见的恶性肿瘤之一，其中HER2基因的状态对于乳腺癌的治疗和预后评估具有重要意义。

荧光原位杂交（FISH）是检测HER2基因状态的重要手段之一。

以下是乳腺癌HER2基因FISH检测判读的标准，主要包括探针信号识别、细胞计数规定、比例阈值判定、非整倍体评估、基因拷贝数判定、结果分类解读以及质量控制要求等方面。

一、探针信号识别在进行FISH检测时，首先要正确识别探针信号。

通常，HER2基因和对照基因（如CEP17）会被标记为不同的颜色。

正常情况下，HER2基因和CEP17的信号应为红色和绿色。

每个细胞核内应有2个CEP17信号和2个HER2信号。

二、细胞计数规定对于每个样本，应至少分析20个非重叠的、完整的、清晰可辨的细胞核。

如果细胞核不完整、重叠或信号模糊，则该细胞核不应纳入计数。

三、比例阈值判定HER2基因与CEP17的比例（HER2/CEP17）是判定HER2基因状态的关键指标。

根据FISH检测的结果，可以将HER2基因状态分为以下几类：1. HER2基因扩增：当HER2/CEP17比例大于2.0时，判定为HER2基因扩增。

2. HER2基因无扩增：当HER2/CEP17比例小于或等于2.0且大于1.8时，判定为HER2基因无扩增。

3. HER2基因不确定：当HER2/CEP17比例小于或等于1.8时，判定为HER2基因状态不确定，需要进一步检测或结合其他检测方法。

四、非整倍体评估在非整倍体细胞中，CEP17信号可能会超过2个或少于2个。

如果样本中存在非整倍体细胞，应将这些细胞排除在计数之外。

五、基因拷贝数判定除了比例阈值外，还可以根据HER2基因和CEP17的拷贝数来判定HER2基因状态。

通常认为，当HER2基因拷贝数大于6时，为HER2基因扩增。

六、结果分类解读根据以上判读标准，可以将FISH检测结果分为以下几类：1. HER2阳性：当HER2基因扩增或基因拷贝数大于6时，判定为HER2阳性。

bass 海鲈鱼,beltfish－带鱼，buffalofish－水牛鱼，carp－鲤鱼catfish 鲶鱼,catfish-鲶鱼，cod 鳕鱼,cod fillets 鳕鱼块cod－鳕鱼，conger (eel)鳗鱼corvina黄花鱼croaker 白花鱼,跟黄花鱼是一类,肉质很嫩,清蒸dace鲮鱼drumfish－鼓鱼flounder 比目鱼,又叫龙利,flounder－龙利, 比目鱼一类的鱼，grouper 石斑鱼haddock 北大西洋鳕鱼halibut ***目鱼herring 青鱼，鲱，herring roes 鲱鱼子mackerel 鲭milkfish－虱目鱼mullet 胭脂鱼orange 香橙鱼octopus 鱆鱼perch 河鲈鱼plaice欧蝶鱼red mullet 红鲣red snapper 红鲷鱼ribbonfish 带鱼.salmon 三文鱼(也叫？鲑鱼？或大马哈鱼sea bream 海鲤shark 鲨鱼swordfish 剑鱼,swordfish－剑鱼，tilapia 罗非鱼,又叫吴郭鱼,非洲鲫鱼,tilapia－鲷鱼，trout－鳟鱼，跟salmon是亲戚tuna－吞拿鱼、金枪鱼，whitefish－白鱼，squid－乌贼，鱿鱼，scallop-扇贝肉，鲤鱼 carp鳗鱼eel章鱼octopus三文鱼salmon鱿鱼 squid金枪鱼 tuna基围虾shrimp虾prawn大虾king prawn龙虾lobster螃蟹crab (crab stick 蟹肉条 )贻贝, 蚌类 mussel牡蛎, 蚝oyster田螺winkleeel 鳗鱼carp 鲤鱼milkfish 遮目鱼salmon 鲑鱼trout 鳟鱼perch 河鲈pomfret 鲳鱼, 银鲳tuna 金枪鱼silver carp 银鲤hairtail 带鱼tilapia 罗非鱼mackerel pike 秋刀鱼sea bream 海鲷conger 海鳗loach泥鳅octopus 章鱼swordfish 旗鱼mullet 胭脂鱼, 鲻鱼abalone 鲍鱼cuttlefish 墨鱼, 乌贼squid 鱿鱼whale 鲸shark 鲨鱼garoupa 石斑goldfish 金鱼guppy 孔雀鱼鲨鱼 shark黄鱼 yellow-fin tuna墨鱼 cuttle鲶鱼 catfish鲫鱼 crucian带鱼 hairtail鲳鱼 pomfret鲈鱼 weever黑鱼 snakehead金鱼 goldfish热带鱼 tropical fish鲫鱼:Carp带鱼: Octopus鲨鱼:Sharks黄花鱼：Croaker墨鱼：Cuttlefish金枪鱼：Tuna鳗鱼： Eel草鱼：Grass carp鲈鱼： Perch鲶鱼： Catfish沙丁鱼：Sardine三文鱼：Salmon大头鱼：Main fish娃娃鱼： Tibetan欢迎您的下载，资料仅供参考！致力为企业和个人提供合同协议，策划案计划书，学习资料等等打造全网一站式需求。