Εισαγωγή στη Ρευστομετρία

Introduction to Flow Cytometry

• Flow cytometry is a laser-based technique for analyzing and sorting individual cells or particles in a liquid suspension.
• It simultaneously measures multiple physical and chemical characteristics of each cell.
• Cells are suspended in a fluid and passed single-file through a laser beam.
• Detectors measure light scatter and fluorescence emitted from the cells.

Key Components of a Flow Cytometer

• Fluidics system: Maintains a consistent stream of cells.
• Laser: Excites fluorescent labels on cells, generating signals for analysis.
• Optical system: Detects light scattered and emitted by the cells.
• Electronics: Amplifies and digitizes the signals.
• Data analysis software: Processes the data and produces results.

Principle of Flow Cytometry

• Cells are labeled with fluorescent markers (e.g., antibodies) that bind to specific cell surface proteins or intracellular components.
• Light scatter: Forward scatter (FSC) indicates cell size; side scatter (SSC) reveals granularity or internal complexity.
• Fluorescence signals: Measured by the intensity of fluorescence emitted by each cell.
• Data is presented in histograms or dot plots, identifying different cell populations based on their characteristics.

Applications of Flow Cytometry

• Cell counting and analysis: Determining cell viability, apoptosis, and cell cycle stage.
• Immune cell phenotyping: Identifying and quantifying various immune cell types.
• Cancer diagnostics and research: Characterizing tumor cells, identifying cancer stem cells, and monitoring treatment response.
• Stem cell research: Analyzing stem cell populations and their differentiation potential.

Data Representation and Analysis

• Histograms: Display cell distribution based on a single parameter's intensity (e.g., fluorescence intensity).
• Dot plots: Show the relationship between two parameters (e.g., SSC vs. FSC or fluorescence vs. fluorescence).
• Gate creation: Allows focusing on specific cell populations based on defined characteristic ranges (e.g., cell size, granularity, fluorescence).
• Statistical analysis: Quantifies the proportions of different cell populations.

Advantages of Flow Cytometry

• High-throughput analysis: Rapid evaluation of numerous cells.
• High resolution: Precise identification and quantification of cell types.
• Multiparametric analysis: Simultaneous measurement of multiple cell characteristics.
• Non-destructive: Cells remain viable for further analysis or culturing after the process.
• High sensitivity: Detects subtle differences in cell populations.

Disadvantages of Flow Cytometry

• Expensive equipment.
• Requires specialized expertise for operation and data analysis.
• Potential for sample loss or damage.
• Fluorescent labels may not always reliably represent the desired target protein or cell type.
• Cell viability issues affect data accuracy and interpretation.

Flow Cytometry in Medical Diagnostics

• Immunophenotyping of leukemias and lymphomas: Differentiates between blood cancer subtypes based on surface marker expression.
• Infectious disease research: Detects and characterizes pathogens or infected cells.
• Monitoring immune cell responses: Evaluates immune responses to vaccines, therapies, or infections.
• Transplant rejection: Studies T cell subset activities to determine transplant success.

Flow Cytometry in Research

• Stem cell research: Examines stem cell differentiation potential under various conditions.

Flow Cytometry Sorting

• Cell sorting: Isolates specific cell populations based on their characteristics using flow cytometry.
• The process involves passing cells through a nozzle, applying an electric charge to target cell populations, which are then directed to different collector tubes.
• This isolates homogeneous cell populations, crucial for numerous experimental purposes.

Workflow & Procedure

• Sample preparation: Cells are suspended in a solution and stained with fluorescent dyes.
• Data acquisition: Cells are passed through the flow cytometer.
• Data analysis: Software analyzes the data, identifying cell populations.
• Sorting (Optional): Specific cell populations are separated from the mixture.

Αυτή η κουίζ διερευνά τις βασικές αρχές της ρευστομετρίας, μια τεχνική ανάλυσης και διάχωσης κυττάρων με χρήση λέιζερ. Εξετάζονται τα κύρια συστατικά ενός ρευστομέτρου και η αρχή λειτουργίας του. Μέσω πολυάριθμων μετρικών, η ρευστομετρία βοηθά στην ανάλυση χαρακτηριστικών κυττάρων και σωματιδίων.