Encyclopedia of Experiments: Biology
A subscription to JoVE is required to view this content. Sign in or start your free trial.
The JoVE video player is compatible with HTML5 and Adobe Flash. Older browsers that do not support HTML5 and the H.264 video codec will still use a Flash-based video player. We recommend downloading the newest version of Flash here, but we support all versions 10 and above.
If that doesn't help, please let us know.
Transcript
Please note that all translations are automatically generated.
Click here for the English version.
- 顕微鏡スライド上の緩衝液の滴から始めて、無傷のワームを液滴に移す。ワームが所定の位置に入ったら、糸くずのない組織を使って緩衝液を浸して取り除き、サンプルを90%エタノールで数回浸してサンプルを固定し、各塗布後に蒸発させる。
エタノールが最終リンスの後に蒸発したら、核酸染料を含む溶液を添加し、DAPIはDNAのマイナーな溝の中でアデニンおよびチミン塩基に優先的に結合する。
次に、アンチフェード保存剤を加えてサンプルの保存期間を延長します。 カバースリップを適用し、スライドにそれをシールします。その後、青色のフィルターを備えた蛍光顕微鏡を使用して、細胞とそのサイクル状態に応じて、単一の染色体、姉妹染色体ペア、または核全体を表すことができるDAPIボディを視覚化します。
- テトラボイド株は12個の染色体を持ち、受精していない卵母細胞にDAPI染色体を数えることで検証できる。そのためには、M9バッファの5〜10マイクロリットルをスライドに落とし、6〜10個の推定四トラボイドをドロップに移す。
その後、10マイクロリットルの90%エタノールをワームに落としてエタノールが完全に蒸発するのを見守る。蒸発が終わったらすぐにエタノールを加えて蒸発するのを見守る。
スライドの長期保存のために、市販のワームまたは自家製のアンチフェードにワームを取り付けます。その後、カバースリップを追加し、マニキュアでエッジを密封します。
まず、最も成熟した未受精卵母細胞を見つけ、 精子に隣接し、まだ精子や子宮のいずれにも入っていない。ここでのDAPI体はおそらく単染色体ペアである。次に、卵母細胞の核に焦点を当て、顕微鏡の細かい焦点を使って上から下までゆっくりとスキャンし、DAPI体を数えながら、同じ核内のDAPI体を数える。
野生型卵母細胞は平均6体のDAPIボディを持つ。12体のDAPIボディの存在は、この株の動物が部分的または完全な四量体であることを示している。
Tags
空の値 発行Related Videos
PLAYLIST
-
Education • BiochemistryPhotometric Protein Determination
-
Education • Basic Methods in Cellular and Molecular BiologySeparating Protein with SDS-PAGE
-
Education • Basic Methods in Cellular and Molecular BiologyThe Western Blot
-
Education • Basic Methods in Cellular and Molecular BiologyThe ELISA Method
-
Education • ImmunologyImmunoprecipitation-Based Techniques: Purification of Endogenous Proteins Using Agarose Beads
-
Education • Basic Methods in Cellular and Molecular BiologyPCR: The Polymerase Chain Reaction
-
Education • Basic Methods in Cellular and Molecular BiologyDNA Gel Electrophoresis
-
Education • Basic Methods in Cellular and Molecular BiologyBacterial Transformation: The Heat Shock Method
-
Education • Basic Methods in Cellular and Molecular BiologyPlasmid Purification
-
Education • Cell BiologyLive Cell Imaging of Mitosis
-
Education • Cell BiologyThe Transwell Migration Assay
-
Education • Cell BiologyCell-surface Biotinylation Assay
-
Education • Cell BiologyFM Dyes in Vesicle Recycling
-
Education • Cell BiologyThe ATP Bioluminescence Assay
-
Education • BiochemistryMetabolic Labeling
-
Education • Cell BiologyThe TUNEL Assay
-
Education • Cell BiologyAnnexin V and Propidium Iodide Labeling
-
Research • BiologyViability Assays for Cells in Culture
-
Education • ImmunologyAssay for Cell Death: Chromium Release Assay of Cytotoxic Ability
-
Education • Basic Methods in Cellular and Molecular BiologyPassaging Cells
-
Education • Basic Methods in Cellular and Molecular BiologyUsing a Hemacytometer to Count Cells
-
Research • BiologyDetermination of Mammalian Cell Counts, Cell Size and Cell Health Using the Moxi Z Mini Automated Cell Counter
-
Education • Basic Methods in Cellular and Molecular BiologyAn Introduction to Transfection
-
Education • NeurosciencePrimary Neuronal Cultures
-
Education • NeuroscienceNeuronal Transfection Methods
-
Research • Developmental BiologyCulture and Transfection of Zebrafish Primary Cells
-
Research • Immunology and InfectionHighly Efficient Transfection of Primary Macrophages with In Vitro Transcribed mRNA
-
Research • BiologyHigh Yield Expression of Recombinant Human Proteins with the Transient Transfection of HEK293 Cells in Suspension
-
Education • Developmental BiologyEmbryonic Stem Cell Culture and Differentiation
-
Education • Developmental BiologyInduced Pluripotency
-
Education • Biology IIBasic Mouse Care and Maintenance
-
Education • Lab Animal ResearchRodent Handling and Restraint Techniques
-
Education • Lab Animal ResearchCompound Administration I
-
Education • Lab Animal ResearchCompound Administration II
-
Education • Lab Animal ResearchCompound Administration III
-
Education • Lab Animal ResearchCompound Administration IV
-
Research • BiologyManual Restraint and Common Compound Administration Routes in Mice and Rats
-
Education • Biology IIIntroducing Experimental Agents into the Mouse
-
Research • BehaviorSystematic Assessment of Well-Being in Mice for Procedures Using General Anesthesia
-
Education • Lab Animal ResearchBlood Withdrawal I
-
Education • Lab Animal ResearchBlood Withdrawal II
-
Education • Lab Animal ResearchAnesthesia Induction and Maintenance
-
Education • Lab Animal ResearchConsiderations for Rodent Surgery
-
Research • NeuroscienceWhole Animal Perfusion Fixation for Rodents
-
Education • Lab Animal ResearchDiagnostic Necropsy and Tissue Harvest
-
Education • General Laboratory TechniquesIntroduction to Light Microscopy
-
Research • BiologyPhase Contrast and Differential Interference Contrast (DIC) Microscopy
-
Education • General Laboratory TechniquesIntroduction to Fluorescence Microscopy
-
Education • Analytical ChemistryScanning Electron Microscopy (SEM)
-
Research • BiochemistryImaging of Extracellular Vesicles by Atomic Force Microscopy
-
Education • ImmunologyConfocal Fluorescence Microscopy: A Technique to Determine the Localization of Proteins in Mouse Fibroblasts
-
Research • BioengineeringVideo-rate Scanning Confocal Microscopy and Microendoscopy
-
Research • BiologyThe CryoAPEX Method for Electron Microscopy Analysis of Membrane Protein Localization Within Ultrastructurally-Preserved Cells
-
Education • General Laboratory TechniquesHistological Sample Preparation for Light Microscopy
-
Education • NeuroscienceHistological Staining of Neural Tissue
-
Research • Encyclopedia of Experiments: BiologyNile Red Staining of C. elegans: A Method to Visualize Lipid Droplets in Fixed Animals
-
Research • Encyclopedia of Experiments: BiologyEthanol Fixation and DAPI Staining: A Method to Visualize DNA in C. elegans
-
Research • NeurosciencePreparation of Non-human Primate Brain Tissue for Pre-embedding Immunohistochemistry and Electron Microscopy
-
Research • Encyclopedia of Experiments: BiologyEmbedding Adult Drosophila Heads in Paraffin: Preparing Tissue for Histology
-
Research • BiologyCryosectioning of Contiguous Regions of a Single Mouse Skeletal Muscle for Gene Expression and Histological Analyses
-
Research • MedicineSampling Strategies and Processing of Biobank Tissue Samples from Porcine Biomedical Models
-
Education • ImmunologyImmunohistochemistry and Immunocytochemistry: Tissue Imaging via Light Microscopy
-
Education • ImmunologyImmunofluorescence Microscopy: Immunofluorescence Staining of Paraffin-Embedded Tissue Sections
-
Research • NeuroscienceImmunostaining of Biocytin-filled and Processed Sections for Neurochemical Markers
-
Education • Developmental BiologyWhole-Mount In Situ Hybridization
-
Education • Core: ChemistryUncertainty in Measurement: Accuracy and Precision
-
Education • Analytical ChemistryInternal Standards
-
Education • Analytical ChemistryCalibration Curves
-
Education • Analytical ChemistryMethod of Standard Addition
-
Education • Analytical ChemistrySample Preparation for Analytical Characterization
-
Education • Analytical ChemistryIntroduction to Mass Spectrometry
-
Education • BiochemistryTandem Mass Spectrometry
-
Research • BioengineeringSample Preparation Strategies for Mass Spectrometry Imaging of 3D Cell Culture Models
-
Research • GeneticsMass Spectrometry-Based Proteomics Analyses Using the OpenProt Database to Unveil Novel Proteins Translated from Non-Canonical Open Reading Frames
-
Education • Materials EngineeringX-ray Diffraction
-
Education • Analytical ChemistryRaman Spectroscopy for Chemical Analysis
-
Education • Analytical ChemistryUltraviolet-Visible (UV-Vis) Spectroscopy
-
Research • EngineeringQuasi-light Storage for Optical Data Packets
-
Education • Analytical ChemistryHigh-Performance Liquid Chromatography (HPLC)
-
Education • Analytical ChemistryGas Chromatography (GC) with Flame-Ionization Detection
-
Education • Biology IIMouse Genotyping
-
Education • GeneticsSNP Genotyping
-
Research • BiologyRapid Genotyping of Mouse Tissue Using Sigma's Extract-N-Amp Tissue PCR Kit
-
Research • BiologyInfinium Assay for Large-scale SNP Genotyping Applications
-
Research • BiologyA PCR-based Genotyping Method to Distinguish Between Wild-type and Ornamental Varieties of Imperata cylindrica
-
Research • Developmental BiologyGenotyping and Quantification of In Situ Hybridization Staining in Zebrafish
-
Research • MedicineCollection and Extraction of Saliva DNA for Next Generation Sequencing
-
Research • BiologyAmplification, Next-generation Sequencing, and Genomic DNA Mapping of Retroviral Integration Sites
-
Research • BiologyNext-generation Sequencing of 16S Ribosomal RNA Gene Amplicons
-
Research • BiologyTargeted DNA Methylation Analysis by Next-generation Sequencing
-
Education • Core: PsychologyMeasures of Central Tendency
-
Education • Core: PsychologyVariation: Normal Distribution, Range, and Standard Deviation
-
Education • Core: PsychologyRegression Toward the Mean
-
Education • Core: PsychologyStatistical Significance
-
Research • NeuroscienceStatistical Modelling of Cortical Connectivity Using Non-invasive Electroencephalograms
-
Research • BiochemistryFully Autonomous Characterization and Data Collection from Crystals of Biological Macromolecules
-
Research • BehaviorAutomated, Quantitative Cognitive/Behavioral Screening of Mice: For Genetics, Pharmacology, Animal Cognition and Undergraduate Instruction
-
Research • Cancer ResearchIntegration of Bioinformatics Approaches and Experimental Validations to Understand the Role of Notch Signaling in Ovarian Cancer
-
Research • BiologyVideo Bioinformatics Analysis of Human Embryonic Stem Cell Colony Growth
-
Research • GeneticsTargeted Next-generation Sequencing and Bioinformatics Pipeline to Evaluate Genetic Determinants of Constitutional Disease
-
Research • GeneticsRNA Next-Generation Sequencing and a Bioinformatics Pipeline to Identify Expressed LINE-1s at the Locus-Specific Level
-
Research • GeneticsA Bioinformatics Pipeline to Accurately and Efficiently Analyze the MicroRNA Transcriptomes in Plants
-
Research • BiologyA Bioinformatics Pipeline for Investigating Molecular Evolution and Gene Expression using RNA-seq