Resources
- FAQ
- Protocol
- Cell Culture Guide
- Technical Bulletins
-
Explore & Learn
-
Cell Biology
- Comparison of Several Techniques for the Detection of Apoptotic Cells
- STR Profiling—The ID Card of Cell Line
- How to Handle Mycoplasma in Cell Culture?
- Enrichment, Isolation and Characterization of Circulating Tumor Cells (CTCs)
- Strategies for Enrichment of Circulating Tumor Cells (CTCs)
- How to Assess the Migratory and Invasive Capacity of Cells?
- Tips For Cell Cryopreservation
- What Cell Lines Are Commonly Used in Biopharmaceutical Production?
- T Cell Activation and Expansion
- How to Isolate PBMCs from Whole Blood?
- CHO Cell Line Development
- How to Isolate and Analyze Tumor-Infiltrating Leukocytes?
- Generation and Applications of Neural Stem Cells
- Stem Cell Markers
- Troubleshooting Cell Culture Contamination: A Comprehensive Guide
- Comparison of the MSCs from Different Sources
- What are the Differences Between M1 and M2 Macrophages?
- Organoid Differentiation from Induced Pluripotent Stem Cells
- Quantification of Cytokines
- Multi-Differentiation of Peripheral Blood Mononuclear Cells
- Mesenchymal Stem Cells: A Comprehensive Exploration
- IL-12 Family Cytokines and Their Immune Functions
- What are Mesothelial Cells?
- How to Scale Up Single-Cell Clones?
- Biomarkers and Signaling Pathways in Tumor Stem Cells
- Techniques for Cell Separation
- Contamination of Cell Cultures & Treatment
- Cell Culture Medium
- What Are Myeloid Cell Markers?
- Cryopreservation of Cells Step by Step
- Cell Cryopreservation Techniques and Practices
- Human Primary Cells: Definition, Assay, Applications
- How to Eliminate Mycoplasma From Cell Cultures?
- Critical Quality Attributes and Assays for Induced Pluripotent Stem Cells
- T Cell, NK Cell Differentiation from Induced Pluripotent Stem Cells
- Major Problems Caused by the Use of Uncharacterized Cell Lines
- Direct vs. Indirect Cell-Based ELISA
- What Is Cell Proliferation and How to Analyze It?
- Unveiling the Molecular Secrets of Adipogenesis in MSCs
- How to Decide Between 2D and 3D Cell Cultures?
- Isolation, Expansion, and Analysis of Natural Killer Cells
- Neural Differentiation from Induced Pluripotent Stem Cells
- Tumor Stem Cells: Identification, Isolation and Therapeutic Interventions
- Guidelines for Cell Banking to Ensure the Safety of Biologics
- Monocytes vs. Macrophages
- How to Detect and Remove Endotoxins in Biologics?
- Comparison of Different Methods to Measure Cell Viability
- What are PBMCs?
- Circulating Tumor Cells as Cancer Biomarkers in the Clinic
- CFU Assay for Hematopoietic Cell
- How to Start Your Culture: Thawing Frozen Cells
- Optimization Strategies of Cell-Based Assays
- Immunogenicity Testing: ELISA and MSD Assays
- 3D-Cell Model in Cell-Based Assay
- Types of Cell Therapy for Cancer
- From Collection to Cure: How ACT Works in Cancer Immunotherapy
- How to Maximize Efficiency in Cell-Based High-Throughput Screening?
- Cell-Based High-Throughput Screening Techniques
- What Are CAR T Cells?
- Live Cell Imaging: Unveiling the Dynamic World of Cellular Processes
- From Blur to Clarity: Solving Resolution Limits in Live Cell Imaging
- From Blur to Clarity: Solving Resolution Limits in Live Cell Imaging
- Live Cell Imaging: Unveiling the Dynamic World of Cellular Processes
- Understanding Immunogenicity Assays: A Comprehensive Guide
- 3D-Cell Model in Cell-Based Assay
- Role of Cell-Based Assays in Drug Discovery and Development
- Immunogenicity Testing: ELISA and MSD Assays
- Optimization Strategies of Cell-Based Assays
- What Are the Pros and Cons of Adoptive Cell Therapy?
- Adherent and Suspension Cell Culture
- Overview of Cell Apoptosis Assays
- Mastering Cell Culture and Cryopreservation: Key Strategies for Optimal Cell Viability and Stability
- Key Techniques in Primary, Immortalized and Stable Cell Line Development
- From Primary to Immortalized: Navigating Key Cell Lines in Biomedical Research
- Exploring Cell Dynamics: Migration, Invasion, Adhesion, Angiogenesis, and EMT Assays
- Cell Viability, Proliferation and Cytotoxicity Assays
-
Histology
- Troubleshooting in Fluorescent Staining
- Instructions for Tumour Tissue Collection, Storage and Dissociation
- Fluorescent Nuclear Staining Dyes
- Multiple Animal Tissue Arrays
- Tips for Choosing the Right Protease Inhibitor
- Guides for Live Cell Imaging Dyes
- Overview of the FFPE Cell Pellet Product Lines
- Immunohistochemistry Troubleshooting
- Cell and Tissue Fixation
- Cell Lysates: Composition, Properties, and Preparation
- Microscope Platforms
- Mitochondrial Staining
- Overview of Common Tracking Labels for MSCs
- How to Apply NGS Technologies to FFPE Tissues?
- Immunohistochemistry Controls
- Stains Used in Histology
- Comparison of Membrane Stains vs. Cell Surface Stains
- Comparing IHC, ICC, and IF: Which One Fits Your Research?
- Common Immunohistochemistry Stains and Their Role in Cancer Diagnosis
- Histological Staining Techniques: From Traditional Chemical Staining to Immunohistochemistry
- From Specimen to Slide: Core Methods in Histological Practice
- Modern Histological Techniques
-
Exosome
- What's the Potential of PELN in Disease Treatment?
- Summary of Approaches for Loading Cargo into Exosomes
- Emerging Technologies and Methodologies for Exosome Research
- Exosomes as Emerging Biomarker Tools for Diseases
- How to Apply Exosomes in Clinical?
- How to Efficiently Utilize MSC Exosomes for Disease Treatment?
- How to characterize exosomes?
- How to Enhancement Exosome Production?
- How to Label Exosomes?
- Classification, Isolation Techniques and Characterization of Exosomes
- How to Perform Targeted Modification of Exosomes?
- How do PELN Deliver Drugs?
- Current Research Status of Milk Exosomes
- Exosome Quality Control: How to Do It?
- The Role of Exosomes in Cancer
- Techniques for Exosome Quantification
- Exosome Size Measurement
- What are the Functions of Exosomal Proteins?
- Applications of MSC-EVs in Immune Regulation and Regeneration
- Unraveling Biogenesis and Composition of Exosomes
- Production of Exosomes: Human Cell Lines and Cultivation Modes
- Exosome Transfection for Altering Biomolecular Delivery
- How Important are Lipids in Exosome Composition and Biogenesis?
- Collection of Exosome Samples and Precautions
- Exosome Antibodies
- Common Techniques for Exosome Nucleic Acid Extraction
-
ISH/FISH
- Reagents Used in FISH Experiments
- Small RNA Detection by ISH Methods
- Telomere Length Measurement Methods
- Comprehensive Comparison of IHC, CISH, and FISH Techniques
- ISH probe labeling method
- What Types of Multicolor FISH Probe Sets Are Available?
- What Is the Use of FISH in Solid Tumors?
- Mapping of Transgenes by FISH
- CARD-FISH: Illuminating Microbial Diversity
- RNAscope ISH Technology
- What are the Differences between FISH, aCGH, and NGS?
- Overview of Oligo-FISH Technology
- Differences Between DNA and RNA Probes
- FISH Tips and Troubleshooting
- Comparative Genomic Hybridization and Its Applications
- In Situ Hybridization Probes
- Multiple Options for Proving Monoclonality
- FISH Techniques for Biofilm Detection
- Whole Chromosome Painting Probes for FISH
- Guidelines for the Design of FISH Probes
- Different Types of FISH Probes for Oncology Research
- How to Use FISH in Hematologic Neoplasms?
- What are Single, Dual, and Multiplex ISH?
- Overview of Common FISH Techniques
- Multiple Approaches to Karyotyping
-
Toxicokinetics & Pharmacokinetics
- Overview of In Vitro Permeability Assays
- What Are Metabolism-Mediated Drug-Drug Interactions?
- How to Improve Drug Plasma Stability?
- Organ-on-a-Chip Systems for Drug Screening
- Experimental Methods for Identifying Drug-Drug Interactions
- Key Considerations in Toxicokinetic
- Pharmacokinetics Considerations for Antibody Drug Conjugates
- Pharmacokinetics of Therapeutic Peptides
- How Is the Cytotoxicity of Drugs Determined?
- How to Improve the Pharmacokinetic Properties of Peptides?
- Toxicokinetics vs. Pharmacokinetics
- Organoids in Drug Discovery: Revolutionizing Therapeutic Research
- Methods of Parallel Artificial Membrane Permeability Assays
- How to Conduct a Bioavailability Assessment?
- Traditional vs. Novel Drug Delivery Methods
- Predictive Modeling of Metabolic Drug Toxicity
- The Rise of In Vitro Testing in Drug Development
- What Are Compartment Models in Pharmacokinetics?
- Comparison of MDCK-MDR1 and Caco-2 Cell-Based Permeability Assays
- What factors influence drug distribution?
- How to Design and Synthesize Antibody Drug Conjugates?
- Key Factors Influencing Brain Distribution of Drugs
- Effects of Cytochrome P450 Metabolism on Drug Interactions
- How to Improve Drug Distribution in the Brain
- Physical and Chemical Properties of Drugs and Calculations
- What Is the Role of the Blood-Brain Barrier in Drug Delivery?
- Parameters of Pharmacokinetics: Absorption, Distribution, Metabolism, Excretion
- What are the Pharmacokinetic Properties of the Antisense Oligonucleotides?
- Unraveling the Role of hERG Channels in Drug Safety
- What Are the Best Methods to Test Cardiotoxicity?
- Why Cardiotoxicity Matters in R&D?
-
Disease Models
- What Human Disease Models Are Available for Drug Development?
- Overview of Cardiovascular Disease Models in Drug Discovery
- Preclinical Models of Acute Liver Failure
- Summary of Advantages and Limitations of Different Oncology Animal Models
- Why Use PDX Models for Cancer Research?
- Disease Models of Diabetes Mellitus
- Animal Models of Neurodegenerative Diseases
-
Cell Biology
- Life Science Articles
- Download Center
- Trending Newsletter
Protocol for the Preparation of Human Peripheral Blood Chromosomes
GUIDELINE
Human peripheral blood small lymphocytes, which are usually in the G1 (or G0) phase, do not normally divide. If phytohemagglutinin (PHA) is added to the culture medium, such small lymphocytes can be stimulated to transform into lymphoblasts and enter mitosis. After short-term culture, a large number of mitotic cells can be obtained by colchicine treatment, hypotonicity, and fixation. 1 ml of human peripheral blood generally contains about 1×106 to 3×106 small lymphocytes, which is sufficient for chromosome specimen preparation and analysis.
METHODS
- Sterilize the skin with alcohol, and collect blood from the elbow vein 0.3-0.5 ml. Immediately pass the syringe needle directly through the rubber stopper of the culture bottle, inject 30-40 drops of whole blood into 10 ml of culture medium, shake gently, and set it in a 37°C constant temperature box for incubation.
- Cultivate for 68 hours. During the incubation period, the cells are shaken gently periodically to make them full contact with the culture medium.
- 2-4 hours before termination of culture, colchicine (final concentration of 0.07 μg/ml) is added to the culture medium.
- Transfer all the cultures into clean centrifuge tubes and centrifuge at 1000 rpm for 8-10 minutes, discard the supernatant.
- Add 8 ml of pre-warmed 37°C hypotonic solution to the graduated centrifuge tube and mix well with a dropper. Place in a 37°C constant temperature water bath for 15-25 minutes.
- After hypotonic, add 0.5 ml of fixative, mix gently, and centrifuge at 1000 rpm for 8-10 minutes.
- Discard the supernatant, add 5 ml of fixative, mix gently, and leave for 20 min. Centrifuge at 1000 rpm and discard the supernatant. Repeat the fixation three times.
- After discarding the supernatant, depending on the number of cells, add the appropriate amount of fixative to make cell suspension.
- Aspirate the cell suspension from a height of 10-20 cm, drop it onto a dry and clean slide, blow gently, and air dry.
- 1:10 Giemsa staining for 5-10 minutes, fine water to wash away excess staining solution, air dry.
- Look for well-dispersed, moderately stained split phases under low magnification, observe chromosome morphology under an oil microscope, and count.
Creative Bioarray Relevant Recommendations
- Peripheral blood mononuclear cells (PBMCs) are cells with a single, round nucleus and are collected from the peripheral or circulating blood by density centrifugation with Ficoll, a polysaccharide. PBMCs include dendritic cells, monocytes, and lymphocytes (T, B, and NK cells).
- Apart from that, Creative Bioarray offers different ranges of human blood samples, such as whole blood, blood serum, plasma, and coagulation factors.
NOTES
Colchicine needs to be sterilized and dispensed. Carnoy fixative and Giemsa stain, need to be prepared at the time of use.
RELATED PRODUCTS & SERVICES
For research use only. Not for any other purpose.