JoVE Encyclopedia of Experiments
Encyclopedia of Experiments: Biology
Encyclopedia of Experiments: Biology
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Transcript
Please note that all translations are automatically generated.
- عند تصور قطرات الدهون، العضيات التخزين داخل الخلايا للدهون المحايدة، تبدأ بإضافة محلول المنظفات، مثل واحد يحتوي على تريتون X-100، إلى العينة.
ثم، أضف النيل الأحمر إلى العينة وحمايته من الضوء، حيث أن الصبغة حساسة للضوء.
عند التعرض للدهون القطبية، مثل الفوسفوليبيدات، يكون طيف انبعاثات النيل الأحمر في الأطوال الموجية الحمراء الأعلى.
لذلك، استخدم القناة الخضراء للمجهر الفلوري، الذي سيجمع انبعاثات الطول الموجي الأصفر والذهبي، لتصور تلطيخ النيل الأحمر لقطرات الدهون في جميع أنحاء الحيوان.
- لتنفيذ تلطيخ الدهون الأحمر النيل، والديدان لوحة على المتوسط نمو النيماتودا، أو NGM، المصنف مع سجل في وقت متأخر OP50 E. القولونية، وتنمو الديدان في 20 درجة مئوية إلى مرحلة L4 في وقت مبكر. غسل الديدان مع 1 ملليلتر من محلول PBST ونقل تعليق الدودة إلى أنبوب ميكروفوج 1.5 ملليلتر.
الطرد المركزي الديدان في 560 مرة الجاذبية لمدة دقيقة واحدة، ثم إزالة supernatant وتكرار غسل PBST حتى يتم مسح القولونية E. من التعليق.
- إضافة الأيزوبروبانول أمر بالغ الأهمية لبيرميليبيليشن السليم من الديدان قبل تلطيخ.
- الطرد المركزي الديدان في 560 مرة الجاذبية لمدة دقيقة واحدة وإزالة supernatant دون تعطيل بيليه دودة.
- من المهم تقليل التعرض للضوء أثناء خطوات الطرد المركزي.
- الآن، بينما في الظلام، إضافة 600 ميكرولتر من محلول العمل NR المعدة سابقا لكل عينة. عكس الأنابيب ثلاث مرات وخلط تماما الديدان والحل.
بعد الحضانة، والطرد المركزي الديدان وإزالة supernatant. ثم إضافة 600 ميكرولتر من PBST واحتضان العينات في الظلام لمدة 30 دقيقة لإزالة وصمة عار NR الزائدة.
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قيمة فارغة، مشكلة ،
PLAYLIST
Molecular biology toolkit
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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
Cell biology toolkit
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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
Basics of cell culture
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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
Basics of animal research
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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
Techniques in microscopy
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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
Techniques in tissue/cell staining
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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
Techniques in analytical chemistry
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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
Techniques in genotyping
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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
Biostatistical analysis, bioinformatics and big data
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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
