Flow Cytometry Kit

Flow Cytometry Kit is a set of reagents and tools used for flow cytometry, a powerful technique for analyzing the physical and chemical characteristics of cells or particles in a fluid stream. This technology enables rapid multiparametric analysis of single cells or particles as they pass through a laser beam.

Technical Content

Principle of Operation

• Flow Cytometry: The technique involves passing cells or particles in a liquid stream through a laser beam. Each cell or particle is analyzed individually based on its size, granularity, and fluorescence emission.
• Fluorescence Detection: Cells or particles are typically labeled with fluorescent dyes or antibodies. The emitted fluorescence is detected and quantified to determine specific characteristics or markers.

Components

• Antibodies/Fluorochromes: Specific antibodies conjugated to fluorescent dyes that bind to target antigens or cellular markers.
• Buffers and Reagents: Solutions used for cell staining, washing, and maintaining cell viability.
• Controls: Positive and negative controls to ensure assay accuracy and validate results.
• Standard Beads: Calibration beads used to set instrument parameters and validate performance.

Procedure

• Sample Preparation: Cells or particles are prepared and stained with fluorescently labeled antibodies or dyes.
• Staining: Incubate cells with the fluorescently labeled antibodies specific to the target markers. This step typically includes a washing phase to remove excess unbound antibodies.
• Flow Cytometry Analysis: The prepared sample is introduced into the flow cytometer, where it is hydrodynamically focused into a single-cell stream.

Parameters Analyzed

• Forward Scatter (FSC): Measures cell size.
• Side Scatter (SSC): Measures cell granularity or internal complexity.
• Fluorescence Intensity: Indicates the presence and quantity of specific markers or molecules.

Sensitivity and Specificity:

• Sensitivity: Flow cytometry provides high sensitivity and can detect rare cell populations or low-abundance markers.
• Specificity: The use of specific antibodies ensures that the assay targets only the desired markers, minimizing non-specific binding.

Applications

• Immunophenotyping: Characterizing cell types based on surface markers in clinical and research settings.
• Cell Cycle Analysis: Assessing the distribution of cells in different phases of the cell cycle.
• Apoptosis Detection: Identifying cells undergoing programmed cell death.
• Quantification of Biomolecules: Measuring intracellular or extracellular molecules in various biological samples.