flow cytometer definition, principal, requirements procedure and application.
- 1. Flow Cytometry Medical lab Instrumentation Group Members: Neha Shahbaz Rimsha Malik Aimen Shabir Tania Saleem Aneeha
- 2. INTRODUCTION: • Flow cytometry is a laser-based technique used to study and analyze the chemical and physical characteristics of cells or particles. • It was first develop in the 1950s and 1960s by Wallace H. Coulter, Mack Fulwyler, and other, and has become an essential tool in many areas of biology, medicine and biotechnology Flow cytometer: • A Flow Cytometer is a specialized laboratory instrument that is used to analyze and sort individual cells based on their physical and Chemical prosperities in a fluid suspension.
- 3. • The basic components of flow cytometer are fluidic system , a laser, a flow cell or cuvette, detector and a Computer
- 5. Principle: The basic principle of flow cytometry is based on the measurement of light scattered by particles, and the fluorescence observed when these particles are passed in a stream through a laser beam. Working principle: • The Flow cytometry work on the principle of Hydrodynamic focusing system • HFS is a technique in which streams of cell or particles are focused into a narrow stream, by using a sheath fluid that surrounded a sample streams and create a laminar flow, which reduce the turbulence and direct the particles into the center of streams. • A laser beam is used to illuminate the cell.
- 6. • Fluorescent dyes or antibody is attached to the cell are excited by the laser and emit light, which is collected by photomultiplier tubed. • The resulting electric signals are proceed and analyzed by the computer, which generate a histogram or scatter plot that display measured parameter of cell.
- 8. • Forward Scattering based on cell size Side Scattering show complexity of cell
- 12. Types of Flow Cytometry: • Analytical Flow Cytometer • These are commonly used type and are designed to measure wide range of cellular properties such as Size, granularity, and fluorescence intensity. • They allow simultaneous detection of multiple fluorescent probe, due to multiple laser lines and detectors. • Sorting Flow Cytometer • These have an additional components called a cell sorts, that allow physical separation if cell based on properties. • The cell sorter uses a series of electrostatic or mechanical device to sort cell into different collection tube or plates. • Imaging Flow Cytometer • These flow cytometer with high-resolution imaging technology. They use specialized cameras to capture high resolution images of individual cells they pass through flow cell allowing foe more detailed analysis of cellular morphology and function.
- 13. • Micro Fluid Cytometer • These flow cytometer use micro fluid channels and chips to analyze and sort cells. • They are often more compact and portable then traditional flow cytometers and can be used for point-of-care diagnostic and other application. • Mass Cytometer • These flow cytometer used a technique called mass cytometry, which measure the mass-to- charge ratio of individual cell rather then their fluorescence. • These allow for the simultaneous analysis of many parameters with high accuracy and resolution and helpful in the study of complex diseases. Applications: • It is used in clinical labs for the detection of malignancy in bodily fluids like leukemia. • Cytometers like cell sorters can be used to separate the cells of interest in separate collection tubes physically. • It can be used for the detection of the content of DNA by using fluorescent markers. • Flow cytometers allow the analysis of replication cells by using fluorescent dye for four different stages of the cell cycle. • A flow cytometers are used in the study of multi-drug resistant bacteria in the blood and other samples.
- 14. Maintaince • Daily Cleaning • Regular Maintenance • Quality Control • Sample separation • Sheath Fluid • Temperature Control