Flow cytometry
•Download as PPTX, PDF•
0 likes•65 views
A fluorescent compound has the property of absorbing light energy at a range of specific wavelengths. This absorption of light causes electron to rise from the ground state to a higher energy level (excited state). The excited electron quickly decays to its ground state while releasing the excess energy in the form of photon of light. This transition of energy is called fluorescence. https://www.creative-bioarray.com/support/fluorochromes-in-flow-cytometry.htm
Report
Share
Report
Share
Recommended
Flow Cytometry
The document provides an overview of the basic principles and components of flow cytometry. It discusses how flow cytometry works by measuring the properties of cells in fluid flow, using a combination of fluidics to introduce cells, optics to generate and collect light signals, and electronics to convert signals to digital data. Key aspects summarized include how cells are hydrodynamically focused and interrogated by light scatter and fluorescence to derive information on their size, granularity, and marker expression that can be analyzed using software.
Flow cytometry
Flow cytometry is a standard laser-based technology that is used in the detection and measurement of physical and chemical characteristics of cells or particles in a heterogeneous fluid mixture.
FACS and MACS with their applications in biological research.
Flow cytometry (FACS) and magnetic activated cell sorting (MACS) are techniques used to analyze and separate cells based on their physical and chemical characteristics. FACS uses lasers to detect cell properties and sort cells into containers one by one, while MACS uses magnetic microbeads attached to cells to separate them in high gradient magnetic fields. These techniques have various applications in research including identifying stem cells, characterizing cancer cells, studying cell cycles, and isolating cell populations for further analysis.
Flow cytometry
Flow cytometry works by passing cells in a fluid stream through a laser beam, which causes light scattering and fluorescence that is detected and analyzed. Cells are labeled with fluorescent markers and passed through the flow cell in a hydrodynamically focused stream. A laser excites the fluorescent molecules, which emit light at different wavelengths. Forward scatter detects cell size, side scatter detects internal complexity. Detectors convert light signals to digital data, which is analyzed through gating and dot plots to identify cell populations and properties. Flow cytometry allows rapid multi-parameter analysis of individual cells.
Flow cytometry
Flow cytometry (FCM) is a technique used to detect and measure physical and chemical characteristics of a population of cells or particles. In this process, a sample containing cells or particles is suspended in a fluid and injected into the flow cytometer instrument.
Introduction to Flow Cytometry
This document provides an introduction to flow cytometry. It defines flow cytometry as the measurement of physical and chemical characteristics of cells as they flow in a fluid stream through a beam of light. It describes the key components of a flow cytometer including fluidics to deliver cells to the laser, optics to excite and collect light, and electronics to amplify and process signals. It explains the different types of signals detected including light scatter and fluorescence, and how these can be used to characterize cells. The document provides guidance on choosing fluorochromes and considerations for multi-color panels such as spectral overlap. It outlines some common applications of flow cytometry and contact details.
Flow cytometry definition, principle, parts, steps, types, uses
Flow cytometry is a technique that uses lasers to detect and measure physical and chemical characteristics of cells or particles in fluid suspension. Cells pass through a laser beam, which scatters light and causes fluorescence that is detected by sensors. Measurements of scattered and fluorescent light provide information about cell size, granularity, and expression of targeted proteins or nucleic acids. Flow cytometry allows rapid multi-parameter analysis of individual cells in heterogeneous populations and is widely used for clinical, research, and industrial applications.
Flow Cytometry
Flow cytometry allows the measurement of physical characteristics of single cells as they flow through a laser beam. It measures size, granularity, and fluorescence. Applications include immunophenotyping cancers and leukemias, monitoring transplant rejection and HIV, and determining DNA content and proliferation. Recent advances include improved instruments, new antibodies, and assessment of cytoplasmic/nuclear antigens and T-cell clonality.
Recommended
Flow Cytometry
The document provides an overview of the basic principles and components of flow cytometry. It discusses how flow cytometry works by measuring the properties of cells in fluid flow, using a combination of fluidics to introduce cells, optics to generate and collect light signals, and electronics to convert signals to digital data. Key aspects summarized include how cells are hydrodynamically focused and interrogated by light scatter and fluorescence to derive information on their size, granularity, and marker expression that can be analyzed using software.
Flow cytometry
Flow cytometry is a standard laser-based technology that is used in the detection and measurement of physical and chemical characteristics of cells or particles in a heterogeneous fluid mixture.
FACS and MACS with their applications in biological research.
Flow cytometry (FACS) and magnetic activated cell sorting (MACS) are techniques used to analyze and separate cells based on their physical and chemical characteristics. FACS uses lasers to detect cell properties and sort cells into containers one by one, while MACS uses magnetic microbeads attached to cells to separate them in high gradient magnetic fields. These techniques have various applications in research including identifying stem cells, characterizing cancer cells, studying cell cycles, and isolating cell populations for further analysis.
Flow cytometry
Flow cytometry works by passing cells in a fluid stream through a laser beam, which causes light scattering and fluorescence that is detected and analyzed. Cells are labeled with fluorescent markers and passed through the flow cell in a hydrodynamically focused stream. A laser excites the fluorescent molecules, which emit light at different wavelengths. Forward scatter detects cell size, side scatter detects internal complexity. Detectors convert light signals to digital data, which is analyzed through gating and dot plots to identify cell populations and properties. Flow cytometry allows rapid multi-parameter analysis of individual cells.
Flow cytometry
Flow cytometry (FCM) is a technique used to detect and measure physical and chemical characteristics of a population of cells or particles. In this process, a sample containing cells or particles is suspended in a fluid and injected into the flow cytometer instrument.
Introduction to Flow Cytometry
This document provides an introduction to flow cytometry. It defines flow cytometry as the measurement of physical and chemical characteristics of cells as they flow in a fluid stream through a beam of light. It describes the key components of a flow cytometer including fluidics to deliver cells to the laser, optics to excite and collect light, and electronics to amplify and process signals. It explains the different types of signals detected including light scatter and fluorescence, and how these can be used to characterize cells. The document provides guidance on choosing fluorochromes and considerations for multi-color panels such as spectral overlap. It outlines some common applications of flow cytometry and contact details.
Flow cytometry definition, principle, parts, steps, types, uses
Flow cytometry is a technique that uses lasers to detect and measure physical and chemical characteristics of cells or particles in fluid suspension. Cells pass through a laser beam, which scatters light and causes fluorescence that is detected by sensors. Measurements of scattered and fluorescent light provide information about cell size, granularity, and expression of targeted proteins or nucleic acids. Flow cytometry allows rapid multi-parameter analysis of individual cells in heterogeneous populations and is widely used for clinical, research, and industrial applications.
Flow Cytometry
Flow cytometry allows the measurement of physical characteristics of single cells as they flow through a laser beam. It measures size, granularity, and fluorescence. Applications include immunophenotyping cancers and leukemias, monitoring transplant rejection and HIV, and determining DNA content and proliferation. Recent advances include improved instruments, new antibodies, and assessment of cytoplasmic/nuclear antigens and T-cell clonality.
FlowCytometry Basics
Flow cytometry is a powerful analytical tool that can analyze up to 10,000 individual cells per second. It works by passing single cells in suspension through a flow cell where they are exposed to a laser. Light scattering and fluorescence emissions from stained cells are then measured using detectors. Flow cytometry has many applications including analyzing cell viability, cell cycle, apoptosis, and cell sorting. It provides benefits like rapid analysis of single cells but also has limitations such as not showing intracellular protein localization and requiring cell preparation.
Imaging Flow Cytometry
1) Flow cytometry works by passing cells in suspension through a laser or light beam which causes them to scatter light. Optical and electronic systems then collect and analyze these light signals.
2) Key components include fluidics to hydrodynamically focus cells, optics to illuminate cells and collect scattered light, and electronics to convert light signals to digital data.
3) Imaging flow cytometry enhances conventional flow cytometry by incorporating microscopy imaging of cells as they flow, allowing analysis of intracellular structures and distributions.
Flow cytometry
Flow cytometry allows for the quantitative and qualitative analysis of cell properties as cells flow in a fluid stream through a laser. Cells are labeled with fluorescent markers and pass through the laser one by one. Light scattering and fluorescence emission are converted to digital signals which provide information on cell size, granularity, and marker expression. Data is displayed as histograms, dot plots, or density plots to identify cell populations and phenotypes.
Flowcytometery
Diagnostic immunology
Definition.
Uses of Flowcytometery.
Instrument Overview.
Principles of flow cytometry.
Light scatter.
Fluorescence.
Gating.
FACS
cell sorter
Flow cytometry
Flow cytometry allows for the quantitative and qualitative analysis of properties of cell populations in suspension. It works by passing cells in single file through a laser, which scatters and fluoresces light detected by sensors. This provides information about cell size, granularity, and fluorescence at a single cell level. It is commonly used to analyze blood, bone marrow, and lymph node samples. Flow cytometry is multiparametric, can rapidly analyze large cell numbers, and detect rare populations. It finds clinical applications in immunophenotyping, cell counting, sorting, and other analyses.
Flow Cytometry Principle
Since its inception, FACS has been used extensively in biomedical research and clinical diagnostics and therapeutics.
Kumcintroductiontoflowcytometry 141106163704-conversion-gate02
Flow cytometry is a technique that allows for the characterization of individual cells as they flow in a fluid stream through a laser beam. Parameters like cell size, granularity, and fluorescence from markers bound to cells can be measured. Key components include hydrodynamic focusing to align cells, lasers to excite fluorochromes, and detectors like photomultiplier tubes to quantify light scatter and emission. Developments in monoclonal antibodies, fluorochromes, and electronics have advanced the field since its origins in the 1930s measuring basic cell properties. Modern flow cytometers precisely route light through filters to characterize thousands of individual cells per second.
Flow cytometry and fluorescence activated cell sorting (FACS)
Flow cytometry is a technology that analyzes physical and chemical characteristics of particles in fluid as they pass through a laser. It is used for cell counting, sorting, biomarker detection, and protein engineering. The basic principle is passing cells in single file past a laser for detection, counting, and sorting. It has applications in leukocyte analysis, DNA analysis, detecting enzymatic deficiencies, minimal residual disease, detecting autoantibodies, fetal-maternal hemorrhage quantification, and reticulocyte analysis.
Flow cytometry
FLOW CYTOMETRY, PRINCIPLE, APPLICATION, USE IN HAEMATOLOGY, COMPONENT OF FLOW CYTOMETRY, DATA INTERPRETATION, DATA ANALYSIS, CELL SHORTING ADVANTAGES AND DISADVANTAGES, IMMUNOLOGICAL CLASSIFICATION OF ACUTE
LEUKEMIA
Kumc introduction to flow cytometry
This document provides an introduction to flow cytometry. It defines flow cytometry as a method for sensing individual cells in a fluid stream as they pass through a laser beam, measuring light scattering and fluorescence. Key aspects of flow cytometry systems and methodology are described, including hydrodynamic focusing of cells, light scattering measurements, use of fluorescent markers, optical and electronic components, data acquisition and analysis techniques like gating and compensation. The history of technological developments in flow cytometry is also summarized.
Cell sorting/FLOW CYTOMETERY/Fluorescence Flow Cytometry
Cell sorting is a technique used to separate specific cell populations from tissues. The first step involves disrupting connections between cells using enzymes or calcium-chelating agents. This converts the tissue into single cells. Flow cytometry can then identify and separate different cell types by measuring light scattering or fluorescence emitted from individually labeled cells as they pass through a laser. Specific cells are labeled with fluorescent antibody markers and can then be isolated from unlabeled cells using a fluorescence activated cell sorter.
Fluorescence- Activated Cell Sorter (FACS)
This document provides an overview of flow cytometry and fluorescence-activated cell sorting (FACS). It describes flow cytometry as a technique for measuring physical and chemical characteristics of cells as they flow in a fluid stream, allowing for single cell analysis. FACS extends this by using fluorescence to identify cell characteristics and sort cells into separate collections based on these characteristics. The key components of a flow cytometer are described as lasers, optics including filters and detectors, fluidics to hydrodynamically focus cells, and electronics to convert optical signals to digital data. Applications including cell phenotyping, apoptosis analysis, and cell cycle analysis are discussed. Cell sorting and quantitative analysis of cell cycle phases are also summarized.
Flow cytometry
Flow cytometry is a technique used in Cell Biology to analyze and measure the volume of cells suspended in a liquid with streamline flow, exposed to a laser beam.
Flow Cytometry technique
All of the information are collected , it's not a research work but I think it will help the students to know about the basic information.
Flow cytometry
Flow cytometry is a technique that uses lasers to illuminate single cell suspensions labeled with fluorescent markers as they flow through the instrument, generating signals from scattered and fluorescent light that can identify cell subsets and characteristics and be analyzed by computer to provide diagnostic information about cell populations. A flow cytometer consists of fluidic, optical, and electronic systems to transport cells to the laser beam, direct resulting light signals to detectors, and convert those signals into electronic data for analysis. Flow cytometry can identify cell types using fluorescent antibodies targeting antigens, analyze DNA content to detect cancer cell abnormalities, and examine the cell cycle to determine proliferation rates of malignant cells.
An introduction to flow cytometry- Ashwini.R
The document provides an introduction to flow cytometry. It describes flow cytometry as a technique that allows simultaneous multiparametric analysis of physical and chemical characteristics of single cells suspended in a fluid stream. Key components of a flow cytometer include fluidics, optics, detectors, and electronics. Cells are hydrodynamically focused into a single file stream and pass through a laser beam, where light scattering and fluorescence emissions provide information about cellular properties. Photodetectors convert light signals into electrical pulses that are analyzed. Flow cytometry has various applications including immunophenotyping, cell sorting, DNA content analysis, and cell cycle/proliferation analysis.
Flow basics2.ppt2
The document provides an overview of flow cytometry, including its history, principles, and applications. It discusses how flow cytometry allows for the measurement of cellular characteristics like fluorescence and light scattering at high speeds. Key developments include the first apparatus for detecting bacteria in a fluid stream in 1947 and the first cell sorter in 1965. The term "fluorescence activated cell sorter" or FACS was coined in 1972. Flow cytometry integrates technologies like lasers, optics, fluidics, and electronics to analyze individual cells and measure parameters such as cell size, granularity, and receptor expression. It has various applications in fields like immunology, genetics, and microbiology.
Flow cytometry for cell componenet analysis
Flow cytometry is a technique that uses lasers and fluorescence to analyze physical and chemical characteristics of cells as they flow in a fluid stream. It allows simultaneous analysis of thousands of cells per second based on parameters like cell size, granularity, and detection of cell surface antigens using specific antibodies labeled with fluorochromes of different colors. Specimens suitable for analysis include blood, bone marrow, body fluids, and cell suspensions generated from tissues. Flow cytometry has various applications like immunophenotyping, DNA analysis, diagnosis of conditions like PNH, reticulated cell counting, and blood bank testing.
Cell Sorting _Flow Cytometry
An introduction to cell sorting by Flow Cytometry. Including a brief overview of the history of droplet based cell sorting as well as how the instruments work. Some information adressing the key criteria to achieving a good cell sort outcome is also included.
Flow Cytometry- Presentation
Flow Cytometry- presentation about the principle, working and applications.
Apt for taking seminars and classes.
flowcytometry-150403122734-conversion-gate01.pdf
Flow cytometry allows for the quantitative and qualitative analysis of cell properties as cells flow in a fluid stream through a laser. Cells are labeled with fluorescent markers and pass through the laser one by one. Light scattering and fluorescence emission are converted to digital signals which can be analyzed as histograms or dot plots. Properties like cell size, granularity, and marker expression are measured. Gating allows analysis of specific cell populations. Flow cytometry has many applications including immunophenotyping, cell cycle analysis, and functional assays.
Flowcytometry by asif
Flow cytometry allows for the quantitative and qualitative analysis of cell populations by measuring physical and chemical characteristics of individual cells as they flow in a single file through a laser beam. Cells are labeled with fluorescent markers and passed through the flow cytometer which measures light scattering and fluorescence emission. Data is collected and analyzed using plots that display fluorescence parameters of individual cells. Flow cytometry has many clinical applications including immunophenotyping, stem cell analysis, and detection of rare cell populations in diseases like cancer.
More Related Content
What's hot
FlowCytometry Basics
Flow cytometry is a powerful analytical tool that can analyze up to 10,000 individual cells per second. It works by passing single cells in suspension through a flow cell where they are exposed to a laser. Light scattering and fluorescence emissions from stained cells are then measured using detectors. Flow cytometry has many applications including analyzing cell viability, cell cycle, apoptosis, and cell sorting. It provides benefits like rapid analysis of single cells but also has limitations such as not showing intracellular protein localization and requiring cell preparation.
Imaging Flow Cytometry
1) Flow cytometry works by passing cells in suspension through a laser or light beam which causes them to scatter light. Optical and electronic systems then collect and analyze these light signals.
2) Key components include fluidics to hydrodynamically focus cells, optics to illuminate cells and collect scattered light, and electronics to convert light signals to digital data.
3) Imaging flow cytometry enhances conventional flow cytometry by incorporating microscopy imaging of cells as they flow, allowing analysis of intracellular structures and distributions.
Flow cytometry
Flow cytometry allows for the quantitative and qualitative analysis of cell properties as cells flow in a fluid stream through a laser. Cells are labeled with fluorescent markers and pass through the laser one by one. Light scattering and fluorescence emission are converted to digital signals which provide information on cell size, granularity, and marker expression. Data is displayed as histograms, dot plots, or density plots to identify cell populations and phenotypes.
Flowcytometery
Diagnostic immunology
Definition.
Uses of Flowcytometery.
Instrument Overview.
Principles of flow cytometry.
Light scatter.
Fluorescence.
Gating.
FACS
cell sorter
Flow cytometry
Flow cytometry allows for the quantitative and qualitative analysis of properties of cell populations in suspension. It works by passing cells in single file through a laser, which scatters and fluoresces light detected by sensors. This provides information about cell size, granularity, and fluorescence at a single cell level. It is commonly used to analyze blood, bone marrow, and lymph node samples. Flow cytometry is multiparametric, can rapidly analyze large cell numbers, and detect rare populations. It finds clinical applications in immunophenotyping, cell counting, sorting, and other analyses.
Flow Cytometry Principle
Since its inception, FACS has been used extensively in biomedical research and clinical diagnostics and therapeutics.
Kumcintroductiontoflowcytometry 141106163704-conversion-gate02
Flow cytometry is a technique that allows for the characterization of individual cells as they flow in a fluid stream through a laser beam. Parameters like cell size, granularity, and fluorescence from markers bound to cells can be measured. Key components include hydrodynamic focusing to align cells, lasers to excite fluorochromes, and detectors like photomultiplier tubes to quantify light scatter and emission. Developments in monoclonal antibodies, fluorochromes, and electronics have advanced the field since its origins in the 1930s measuring basic cell properties. Modern flow cytometers precisely route light through filters to characterize thousands of individual cells per second.
Flow cytometry and fluorescence activated cell sorting (FACS)
Flow cytometry is a technology that analyzes physical and chemical characteristics of particles in fluid as they pass through a laser. It is used for cell counting, sorting, biomarker detection, and protein engineering. The basic principle is passing cells in single file past a laser for detection, counting, and sorting. It has applications in leukocyte analysis, DNA analysis, detecting enzymatic deficiencies, minimal residual disease, detecting autoantibodies, fetal-maternal hemorrhage quantification, and reticulocyte analysis.
Flow cytometry
FLOW CYTOMETRY, PRINCIPLE, APPLICATION, USE IN HAEMATOLOGY, COMPONENT OF FLOW CYTOMETRY, DATA INTERPRETATION, DATA ANALYSIS, CELL SHORTING ADVANTAGES AND DISADVANTAGES, IMMUNOLOGICAL CLASSIFICATION OF ACUTE
LEUKEMIA
Kumc introduction to flow cytometry
This document provides an introduction to flow cytometry. It defines flow cytometry as a method for sensing individual cells in a fluid stream as they pass through a laser beam, measuring light scattering and fluorescence. Key aspects of flow cytometry systems and methodology are described, including hydrodynamic focusing of cells, light scattering measurements, use of fluorescent markers, optical and electronic components, data acquisition and analysis techniques like gating and compensation. The history of technological developments in flow cytometry is also summarized.
Cell sorting/FLOW CYTOMETERY/Fluorescence Flow Cytometry
Cell sorting is a technique used to separate specific cell populations from tissues. The first step involves disrupting connections between cells using enzymes or calcium-chelating agents. This converts the tissue into single cells. Flow cytometry can then identify and separate different cell types by measuring light scattering or fluorescence emitted from individually labeled cells as they pass through a laser. Specific cells are labeled with fluorescent antibody markers and can then be isolated from unlabeled cells using a fluorescence activated cell sorter.
Fluorescence- Activated Cell Sorter (FACS)
This document provides an overview of flow cytometry and fluorescence-activated cell sorting (FACS). It describes flow cytometry as a technique for measuring physical and chemical characteristics of cells as they flow in a fluid stream, allowing for single cell analysis. FACS extends this by using fluorescence to identify cell characteristics and sort cells into separate collections based on these characteristics. The key components of a flow cytometer are described as lasers, optics including filters and detectors, fluidics to hydrodynamically focus cells, and electronics to convert optical signals to digital data. Applications including cell phenotyping, apoptosis analysis, and cell cycle analysis are discussed. Cell sorting and quantitative analysis of cell cycle phases are also summarized.
Flow cytometry
Flow cytometry is a technique used in Cell Biology to analyze and measure the volume of cells suspended in a liquid with streamline flow, exposed to a laser beam.
Flow Cytometry technique
All of the information are collected , it's not a research work but I think it will help the students to know about the basic information.
Flow cytometry
Flow cytometry is a technique that uses lasers to illuminate single cell suspensions labeled with fluorescent markers as they flow through the instrument, generating signals from scattered and fluorescent light that can identify cell subsets and characteristics and be analyzed by computer to provide diagnostic information about cell populations. A flow cytometer consists of fluidic, optical, and electronic systems to transport cells to the laser beam, direct resulting light signals to detectors, and convert those signals into electronic data for analysis. Flow cytometry can identify cell types using fluorescent antibodies targeting antigens, analyze DNA content to detect cancer cell abnormalities, and examine the cell cycle to determine proliferation rates of malignant cells.
An introduction to flow cytometry- Ashwini.R
The document provides an introduction to flow cytometry. It describes flow cytometry as a technique that allows simultaneous multiparametric analysis of physical and chemical characteristics of single cells suspended in a fluid stream. Key components of a flow cytometer include fluidics, optics, detectors, and electronics. Cells are hydrodynamically focused into a single file stream and pass through a laser beam, where light scattering and fluorescence emissions provide information about cellular properties. Photodetectors convert light signals into electrical pulses that are analyzed. Flow cytometry has various applications including immunophenotyping, cell sorting, DNA content analysis, and cell cycle/proliferation analysis.
Flow basics2.ppt2
The document provides an overview of flow cytometry, including its history, principles, and applications. It discusses how flow cytometry allows for the measurement of cellular characteristics like fluorescence and light scattering at high speeds. Key developments include the first apparatus for detecting bacteria in a fluid stream in 1947 and the first cell sorter in 1965. The term "fluorescence activated cell sorter" or FACS was coined in 1972. Flow cytometry integrates technologies like lasers, optics, fluidics, and electronics to analyze individual cells and measure parameters such as cell size, granularity, and receptor expression. It has various applications in fields like immunology, genetics, and microbiology.
Flow cytometry for cell componenet analysis
Flow cytometry is a technique that uses lasers and fluorescence to analyze physical and chemical characteristics of cells as they flow in a fluid stream. It allows simultaneous analysis of thousands of cells per second based on parameters like cell size, granularity, and detection of cell surface antigens using specific antibodies labeled with fluorochromes of different colors. Specimens suitable for analysis include blood, bone marrow, body fluids, and cell suspensions generated from tissues. Flow cytometry has various applications like immunophenotyping, DNA analysis, diagnosis of conditions like PNH, reticulated cell counting, and blood bank testing.
Cell Sorting _Flow Cytometry
An introduction to cell sorting by Flow Cytometry. Including a brief overview of the history of droplet based cell sorting as well as how the instruments work. Some information adressing the key criteria to achieving a good cell sort outcome is also included.
Flow Cytometry- Presentation
Flow Cytometry- presentation about the principle, working and applications.
Apt for taking seminars and classes.
What's hot (20)
Kumcintroductiontoflowcytometry 141106163704-conversion-gate02
Kumcintroductiontoflowcytometry 141106163704-conversion-gate02
Flow cytometry and fluorescence activated cell sorting (FACS)
Flow cytometry and fluorescence activated cell sorting (FACS)
Cell sorting/FLOW CYTOMETERY/Fluorescence Flow Cytometry
Cell sorting/FLOW CYTOMETERY/Fluorescence Flow Cytometry
Similar to Flow cytometry
flowcytometry-150403122734-conversion-gate01.pdf
Flow cytometry allows for the quantitative and qualitative analysis of cell properties as cells flow in a fluid stream through a laser. Cells are labeled with fluorescent markers and pass through the laser one by one. Light scattering and fluorescence emission are converted to digital signals which can be analyzed as histograms or dot plots. Properties like cell size, granularity, and marker expression are measured. Gating allows analysis of specific cell populations. Flow cytometry has many applications including immunophenotyping, cell cycle analysis, and functional assays.
Flowcytometry by asif
Flow cytometry allows for the quantitative and qualitative analysis of cell populations by measuring physical and chemical characteristics of individual cells as they flow in a single file through a laser beam. Cells are labeled with fluorescent markers and passed through the flow cytometer which measures light scattering and fluorescence emission. Data is collected and analyzed using plots that display fluorescence parameters of individual cells. Flow cytometry has many clinical applications including immunophenotyping, stem cell analysis, and detection of rare cell populations in diseases like cancer.
Flow cytometry principle
Fluorescent activated cell sorting (FACS) is a specialized type of flow cytometry used for sorting and analyzing a heterogeneous mixture of cells into different sub- populations based on the specific light scattering and fluorescent characteristics (from the specific labels) of each cell.
Flow cytometry-converted biology
Flow cytometry is a technique that uses lasers and fluidics to analyze cells suspended in a fluid stream. Cells pass through a laser beam one by one, scattering and fluorescing light in specific ways depending on their properties. Detectors then measure these light signals, allowing the system to characterize physical and biochemical features of thousands of cells rapidly. Flow cytometry can not only analyze cells but also sort live cells into collections based on their detected characteristics using electrically charged droplets.
AUTOMATION IN HEMATOLOGY
The document discusses automation in hematology. It describes how Wallace Coulter invented the first automated cell counter using electrical impedance to count and size cells. Automation provides advantages like speed, accuracy, and reduced labor but also has disadvantages like erroneous results. There are semi-automated and fully automated analyzers that use various principles like electrical impedance, light scatter, fluorescence, and electrical conductivity to measure cell parameters and provide diagnostic information. Modern analyzers can perform complete blood counts and immunophenotyping to aid in diagnosing conditions like leukemia.
FLOWCYTOMETRY........................pptx
Flow cytometry allows for the quantitative and qualitative analysis of physical and chemical characteristics of cells as they flow in a fluid stream through a laser beam. It measures properties like cell size, granularity, and fluorescence. The main components are the fluidic system that orders cells into a single-file stream, the optical system that applies light and collects signals, and the electronic system that converts signals to digital data. As cells pass through the laser, they scatter and fluoresce light that is detected and analyzed to characterize cell populations and identify abnormal cells. It is used for applications like leukemia diagnosis and monitoring transplant engraftment.
Flow Cytometry for sorting of cells.pptx
Flow cytometry is a technique that uses lasers to detect and measure physical and chemical characteristics of cells or particles as they flow in a fluid stream through a beam of light. Cells are analyzed at rates of thousands per second. Measurements of light scattering and fluorescence are used to differentiate cell types and identify properties like size, granularity, and marker expression. Flow cytometry is used for applications like cell counting, sorting, analyzing function and characteristics, and diagnosing blood cancers.
Hematopoetic Stem Cell enumeration by Flowcytometry
Its about overall procedure,application and gating technique of flowcytometry in Stem cell enumeration
Flow cytometry
This document provides an overview of flow cytometry, including its history, principles, components, applications, and quality control. Flow cytometry involves measuring physical and chemical properties of cells or particles as they pass through a fluid stream. Key developments included Moldavan's early work in 1934 and the coinage of the term "flow cytometry" in the mid-1970s. The three main systems of a flow cytometer are fluidics to transport particles, optics like lasers and detectors, and electronics to convert light signals to data. Applications include clinical uses like detection of bacteria and characterization of cells and particles across many fields.
Flow cytometry and fluorescence microscopy
Flow Cytometry (FC) determines multiple physical and biological characteristics of cells by detecting the intensity of the scattered or emitted light of a single cell in a linear flow state before laser irradiation. This technology can analyze cells rapidly at a rate of tens of thousands of cells per second. The types of samples can be various types of cells (such as peripheral blood, bone marrow, solid tissues, cells in suspension or adherent culture), microorganisms, synthetic microspheres, etc. https://www.creative-proteomics.com/services/flow-cytometry-facs-service.htm
flow cytometry by aksharaditya shukla.
Flow cytometry measures properties of cells in a fluid stream by staining cells with fluorescent antibodies and analyzing them with a flow cytometer instrument. It allows for the characterization of cell populations based on cell surface and intracellular markers. Flow cytometry is useful for immunophenotyping in diseases like leukemia where it can identify the cell lineage involved (lymphoid vs myeloid) and detect minimal residual disease during treatment. In this case, the 5-year old boy presenting with fever, flow cytometry immunophenotyping should be performed to diagnose causes like acute lymphoblastic leukemia by examining markers like CD19, CD5, CD10 on lymphocytes.
Flow cytometry
Flow cytometry allows for the quantitative and qualitative analysis of physical and chemical characteristics of cells and other particles. It works by passing cells in single file through a laser beam, where they scatter and fluoresce light. Detectors then measure these light signals to determine characteristics like cell size, granularity, and marker expression. Key components include a flow cell that hydrodynamically focuses cells, lasers and optical filters to excite and collect signals, and electronics to analyze resulting data. Flow cytometry finds applications in areas like immunophenotyping, disease diagnosis and monitoring treatment response.
Fluorescence-activated Cell Sorting (FACS).pptx
Fluorescence-activated cell sorting (FACS) is a specialized type of flow cytometry that allows for the sorting of cells into containers one by one based on their light scattering and fluorescent characteristics. FACS works by labeling cells with fluorescent markers and passing them through a laser beam, where detectors measure the light scattering and fluorescence to identify cell properties. This technique allows researchers to isolate targeted cell groups for further analysis and is commonly used in medical research areas like hematology and oncology.
Microbiological techniques
This document discusses several common microbiological techniques used to study microbes. It describes microscopy techniques like brightfield and phase contrast microscopy used to view microbes. It also discusses gel electrophoresis to separate DNA, RNA, and proteins by size and charge. Flow cytometry is described as a method to analyze physical and chemical properties of particles using fluorescence. Additional techniques covered include cell counting, spectrophotometry, plating, and polymerase chain reaction (PCR) to amplify DNA segments.
Flow Cytometry (FCM)
Measuring properties of cell (sample) as they flow in a fluid suspension across an illuminated light path.
Presentation on flow cytometry1
Flow cytometry allows for multiparametric analysis of physical and chemical characteristics of particles like cells. It works by passing particles in a fluid stream through a laser beam, detecting light scattering and fluorescence. Main applications include diagnosis of hematological malignancies through immunophenotyping, analysis of DNA content and cell cycle, and organ transplant monitoring. New developments aim to improve sensitivity and integrate imaging capabilities.
Flow cytometry
This document provides an overview of flow cytometry. It begins by defining flow cytometry as a technique for quantitative single cell analysis that counts, examines, and sorts cells based on optical properties like light scattering and fluorescence. It then describes the basic principles, components, and working of a flow cytometer. Key components include lasers, optical filters, and detectors. Cells in suspension pass through the laser one by one, with signals detected by photodiodes and photomultiplier tubes. Applications discussed include cell sorting, apoptosis analysis using markers like Annexin V and PI, and cell cycle analysis using DNA binding dyes or BrdU incorporation. Clinical uses involve hematologic malignancy diagnosis, residual disease detection, and monitoring treatments.
Flow cytometry by kk sahu
Introduction
Definition
Basic mechanism
Prerequisite of flow cytometer
Components of flow cytometry
Flow system
Optics system
Concept of scattering
Advantage
Limitation
Application
Conclusion
References
flow cytometry presentation
This document summarizes flow cytometry, a technique used to count and examine microscopic particles suspended in fluid. It describes key components of modern flow cytometers including lasers, detectors, and computer systems that can analyze thousands of particles per second. The document outlines the principles of how each cell passes through a laser, scatters and emits light, which is detected and analyzed by software. Common applications like cell sorting, fluorescence detection using labeled antibodies, and measurable parameters are discussed. Terminology related to instrumentation, optical systems, data analysis and compensation are also introduced.
Flow cytometry in cell biology
Flow cytometry is a technique used in cell biology that allows for the analysis of physical and chemical characteristics of cells as they flow in a fluid stream through a beam of light. It provides rapid multi-parametric analysis of cells based on light scattering, fluorescence, and other optical properties. Flow cytometry gives information about cell size, granularity, and the expression of cell surface markers or intracellular proteins through the use of fluorescent probes. It has many applications in fields like immunology, cancer research, and infectious disease.
Similar to Flow cytometry (20)
Hematopoetic Stem Cell enumeration by Flowcytometry
Hematopoetic Stem Cell enumeration by Flowcytometry
More from Creative-Bioarray
Kinase Screening
Creative Bioarray offers various kinase assay technologies to support drug discovery research objectives. These include radiometric assays, which are the gold standard but labor intensive, and fluorescence-based assays that are better suited for high-throughput screening. Each technology has strengths and weaknesses depending on the research goal, and Creative Bioarray can support customers through selecting the appropriate assay and interpreting results.
RNA ISH ASSAY
Creative Bioarray provides an extensive range of high quality RNA samples which are ideal for Northern blotting, ribonuclease protection assay, SI nuclease assay, RT-PCR/Q-PCR analysis, rapid amplification of cDNA ends (RACE) and purification of mRNA for library construction.
https://www.creative-bioarray.com/products/rna-list-22.htm
Immunohistochemistry
Immunohistochemistry uses antibodies to detect antigens in tissue samples in order to localize proteins and study biological processes. It has various applications including cancer prognosis, tumor identification, and research. The process involves deparaffinizing tissue, antigen retrieval, blocking, primary/secondary antibody incubation, signal development using enzymes, counterstaining, and mounting slides. Different methods like direct, indirect, PAP, ABC, and LSAB are used depending on the desired sensitivity and specificity. Careful antibody selection and protocol optimization are important for obtaining high quality results.
Flow cytometry
A fluorescent compound has the property of absorbing light energy at a range of specific wavelengths. This absorption of light causes electron to rise from the ground state to a higher energy level (excited state). The excited electron quickly decays to its ground state while releasing the excess energy in the form of photon of light. This transition of energy is called fluorescence.
https://www.creative-bioarray.com/support/fluorochromes-in-flow-cytometry.htm
Caco-2 Permeability Assay
Caco-2 cells are the most widely used cell line for predicting intestinal drug absorption as they form polarized monolayers with tight junctions and microvilli like intestinal cells. Other cell lines used include MDCK, HT29-MTX, 2/4/A1, TC-7, LLC-PK1, IEC-18 and T84, but they have limitations such as lack of transporters, unstable monolayers, or poor differentiation. While cell models show good correlation with passive drug permeability, correlations with active transport are more variable. Extensive studies are still needed to fully characterize intestinal drug transport mechanisms in these models.
Comparison of caco 2 with other cell-based models for intestinal permeability...
The use of cell cultures provides a method to predict drug permeability by utilizing cell monolayers in a two-chamber diffusion system to simulate the passage of drugs from the intestinal lumen into the blood.
Cell cycle regulation
Cell cycle refers to the set of events through which a cell grows, replicates its genome, and ultimately divides into two daughter cells through the process of mitosis.
https://www.creative-bioarray.com/cell-cycle-assays.htm
Caco 2 cell permeability assay
Creative Bioarray is offering Caco-2 permeability assay to help determine the absorption and the bioavailability of drug candidates, facilitating the lead optimization process in drug discovery.
https://www.creative-bioarray.com/Services/caco-2-permeability-assay.htm
Technologies to study kinases
In recent years, several different types of kinase assay technology have been developed, each of which have their relative merits and drawbacks.
Differentiation of ipsc
A broad range of reprogramming systems is available including mRNA reprogramming, dox-inducible human 4F2A reprogramming and lentivirus reprogramming. These technologies have been adopted and developed in close collaboration with leading iPS cell pioneers. Our extensive range of products allows for customization of individual transcription factors, cell culture conditions, and delivery systems for the optimization of reprogramming parameters.
https://www.creative-bioarray.com/Services/Custom-iPS-Cell-Services.htm
Cell Viability
Cell proliferation assays are used to monitor the dynamic growth of a cell population or to detect daughter cells in a growing population.
On the other hand, cell viability assays assess how healthy the cells are by measuring markers of cellular activity.
https://www.creative-bioarray.com/products/cell-viability,-proliferation-and-cytotoxicity-list-226.htm
Cell cycle regulation
Cell cycle regulation is controlled by cyclins, cyclin-dependent kinases (CDKs), and CDK inhibitors. Cyclins bind to and activate CDKs to promote cell cycle progression. CDK inhibitors like p16 and p21 inhibit CDK activity to induce cell cycle arrest. Checkpoints like the DNA damage checkpoint ensure DNA integrity before replication or division. Dysregulation of these regulators can lead to uncontrolled cell growth and cancer.
Cell cycle regulation
By using flow cytometry, staining dyes are needed. Creative Bioarray can choose different dyes to perform the assays, including propidium iodide (PI), BrdU, 7-amino actinomycin-D (7-AAD), Hoechst 33342 and 33258, and 4’6’-diamidino-2-phenylindole (DAPI), based on the customer’s applications or requirements.
https://www.creative-bioarray.com/cell-cycle-assays.htm
Spotlight on ipsc reprogramming
Induced Pluripotent Stem Cells (iPSCs) are a type pf pluripotent stem cell artificially derived, and often referred to as programmed, from adult somatic cells using the expression of certain genes in culture.
https://www.creative-bioarray.com/products/ipsc-reprogramming-kit-list-239.htm
Histology services
Histology Services provides various histology and microscopy services including immunohistochemistry, immunofluorescence, fluorescent in situ hybridization, transmission electron microscopy, and other services to help scientists with life science research. Their services offer flexible and customized solutions to help clients reach research milestones faster and more efficiently while saving on infrastructure costs. They have established proprietary protocols and techniques to enhance sensitivity and quality for applications such as cell and tissue sample preparation, probe design and labeling, hybridization, and imaging.
Primary cells
Creative Bioarray offers 35 human cell systems with over 160 different cell types. Moreover, we also provide our customers with primary cells from over 13 types of other animals.
https://www.creative-bioarray.com/products/primary-cell-types-list-14.htm?page=1
Atp binding cassette (abc) transporter
Drug transporters mediate the uptake and efflux of a broad variety of drugs and drug metabolites. Most uptake transporters are members of the solute carrier (SLC) family, while most efflux transporters are members of the ATP-binding cassette (ABC) transporter family.
https://www.creative-bioarray.com/services/drug-transporters.htm
iPSC-derived Cardiomyocytes
Acroscell provides ready-to-use beating human induced pluripotent stem cells (iPSC)-derived cardiomyocytes. Generated from mature cells that have been genetically reprogramed to a pluripotent stem cell state, induced pluripotent stem cells (iPSCs) can be readily expanded and induced to specialize or differentiate into cardiomyocytes in vitro.
https://www.creative-bioarray.com/acroscell/ipsc-derived-cardiomyocytes.html
PAMPA permeability assay
PAMPA provides a simplified approach to permeability by addressing just a single transport mechanism. This avoids the complexities of active transport and metabolism, enabling ranking of the compounds on a single permeability factor.
https://dda.creative-bioarray.com/parallel-artificial-membrane-permeability-assay.html
Angiogenesis
Creative Bioarray's SuperQuick® angiogenesis assay kit provides a robust method to determine angiogenesis (in vitro) in less than 18 hrs. This assay kit provides a simple, easy to perform, semi-quantitative tool for assessing angiogenesis.
https://www.creative-bioarray.com/superquick-angiogenesis-assay-kit-item-csk-xa001-5627.htm
More from Creative-Bioarray (20)
Comparison of caco 2 with other cell-based models for intestinal permeability...
Comparison of caco 2 with other cell-based models for intestinal permeability...
Recently uploaded
Unlocking the Secrets to Safe Patient Handling.pdf
Furthermore, the time constraints and workload in healthcare settings can make it challenging for caregivers to prioritise safe patient handling Australia practices, leading to shortcuts and increased risks.
MBC Support Group for Black Women – Insights in Genetic Testing.pdf
Christina Spears, breast cancer genetic counselor at the Ohio State University Comprehensive Cancer Center, joined us for the MBC Support Group for Black Women to discuss the importance of genetic testing in communities of color and answer pressing questions.
Champions of Health Spotlight On Leaders Shaping Germany's Healthcare.pdf
This edition features a handful of Champions of Health: Spotlight On Leaders Shaping Germany's Healthcare that are leading us into a better future.
CCSN_June_06 2024_jones. Cancer Rehabpptx
About this webinar: This talk will introduce what cancer rehabilitation is, where it fits into the cancer trajectory, and who can benefit from it. In addition, the current landscape of cancer rehabilitation in Canada will be discussed and the need for advocacy to increase access to this essential component of cancer care.
Let's Talk About It: Breast Cancer (What is Mindset and Does it Really Matter?)
Your mindset is the way you make sense of the world around you. This lens influences the way you think, the way you feel, and how you might behave in certain situations. Let's talk about mindset myths that can get us into trouble and ways to cultivate a mindset to support your cancer survivorship in authentic ways. Let’s Talk About It!
Empowering ACOs: Leveraging Quality Management Tools for MIPS and Beyond
Join us as we delve into the crucial realm of quality reporting for MSSP (Medicare Shared Savings Program) Accountable Care Organizations (ACOs).
In this session, we will explore how a robust quality management solution can empower your organization to meet regulatory requirements and improve processes for MIPS reporting and internal quality programs. Learn how our MeasureAble application enables compliance and fosters continuous improvement.
Can Allopathy and Homeopathy Be Used Together in India.pdf
This article explores the potential for combining allopathy and homeopathy in India, examining the benefits, challenges, and the emerging field of integrative medicine.
CANSA support - Caring for Cancer Patients' Caregivers
International Cancer Survivors Day is celebrated during June, placing the spotlight not only on cancer survivors, but also their caregivers.
CANSA has compiled a list of tips and guidelines of support:
https://cansa.org.za/who-cares-for-cancer-patients-caregivers/
Professional Secrecy: Forensic Medicine Lecture
Professional Secrecy: Forensic Medicine Lecture , Medical jurisprudence
Deep Leg Vein Thrombosis (DVT): Meaning, Causes, Symptoms, Treatment, and Mor...
Deep Leg Vein Thrombosis (DVT): Meaning, Causes, Symptoms, Treatment, and Mor...The Lifesciences Magazine
Deep Leg Vein Thrombosis occurs when a blood clot forms in one or more of the deep veins in the legs. These clots can impede blood flow, leading to severe complications.Dr. David Greene R3 stem cell Breakthroughs: Stem Cell Therapy in Cardiology
Dr. David Greene, founder and CEO of R3 Stem Cell, is at the forefront of groundbreaking research in the field of cardiology, focusing on the transformative potential of stem cell therapy. His latest work emphasizes innovative approaches to treating heart disease, aiming to repair damaged heart tissue and improve heart function through the use of advanced stem cell techniques. This research promises not only to enhance the quality of life for patients with chronic heart conditions but also to pave the way for new, more effective treatments. Dr. Greene's work is notable for its focus on safety, efficacy, and the potential to significantly reduce the need for invasive surgeries and long-term medication, positioning stem cell therapy as a key player in the future of cardiac care.
Michigan HealthTech Market Map 2024 with Policy Makers, Academic Innovation C...
Michigan HealthTech Market Map 2024. Includes 7 categories: Policy Makers, Academic Innovation Centers, Digital Health Providers, Healthcare Providers, Payers / Insurance, Device Companies, Life Science Companies, Innovation Accelerators. Developed by the Michigan-Israel Business Accelerator
Innovative Minds France's Most Impactful Healthcare Leaders.pdf
This edition features a handful of Innovative Minds: France's Most Impactful Healthcare Leaders that are leading us into a better future.
RECENT ADVANCES IN BREAST CANCER RADIOTHERAPY
ULTRA HYPOFRACTIONATION
Accelerated Partial Breast Irradiation: :Brachytherapy
UK FAST FORWARD
Hypotension and role of physiotherapy in it
This particular slides consist of- what is hypotension,what are it's causes and it's effect on body, risk factors, symptoms,complications, diagnosis and role of physiotherapy in it.
This slide is very helpful for physiotherapy students and also for other medical and healthcare students.
Here is the summary of hypotension:
Hypotension, or low blood pressure, is when the pressure of blood circulating in the body is lower than normal or expected. It's only a problem if it negatively impacts the body and causes symptoms. Normal blood pressure is usually between 90/60 mmHg and 120/80 mmHg, but pressures below 90/60 are generally considered hypotensive.
Gemma Wean- Nutritional solution for Artemia
GEMMA Wean is a high end larval co-feeding and weaning diet aimed at Artemia optimisation and is fortified with a high level of proteins and phospholipids. GEMMA Wean provides the early weaned juveniles with dedicated fish nutrition and is an ideal follow on from GEMMA Micro or Artemia.
GEMMA Wean has an optimised nutritional balance and physical quality so that it flows more freely and spreads readily on the water surface. The balance of phospholipid classes to- gether with the production technology based on a low temperature extrusion process improve the physical aspect of the pellets while still retaining the high phospholipid content.
GEMMA Wean is available in 0.1mm, 0.2mm and 0.3mm. There is also a 0.5mm micro-pellet, GEMMA Wean Diamond, which covers the early nursery stage from post-weaning to pre-growing.
Recently uploaded (20)
Unlocking the Secrets to Safe Patient Handling.pdf
Unlocking the Secrets to Safe Patient Handling.pdf
Bringing AI into a Mid-Sized Company: A structured Approach
Bringing AI into a Mid-Sized Company: A structured Approach
MBC Support Group for Black Women – Insights in Genetic Testing.pdf
MBC Support Group for Black Women – Insights in Genetic Testing.pdf
Champions of Health Spotlight On Leaders Shaping Germany's Healthcare.pdf
Champions of Health Spotlight On Leaders Shaping Germany's Healthcare.pdf
Let's Talk About It: Breast Cancer (What is Mindset and Does it Really Matter?)
Let's Talk About It: Breast Cancer (What is Mindset and Does it Really Matter?)
Empowering ACOs: Leveraging Quality Management Tools for MIPS and Beyond
Empowering ACOs: Leveraging Quality Management Tools for MIPS and Beyond
Can Allopathy and Homeopathy Be Used Together in India.pdf
Can Allopathy and Homeopathy Be Used Together in India.pdf
CANSA support - Caring for Cancer Patients' Caregivers
CANSA support - Caring for Cancer Patients' Caregivers
Deep Leg Vein Thrombosis (DVT): Meaning, Causes, Symptoms, Treatment, and Mor...
Deep Leg Vein Thrombosis (DVT): Meaning, Causes, Symptoms, Treatment, and Mor...
Dr. David Greene R3 stem cell Breakthroughs: Stem Cell Therapy in Cardiology
Dr. David Greene R3 stem cell Breakthroughs: Stem Cell Therapy in Cardiology
Michigan HealthTech Market Map 2024 with Policy Makers, Academic Innovation C...
Michigan HealthTech Market Map 2024 with Policy Makers, Academic Innovation C...
Innovative Minds France's Most Impactful Healthcare Leaders.pdf
Innovative Minds France's Most Impactful Healthcare Leaders.pdf
Flow cytometry
- 2. Flow =the motion characteristics of fluids Cytometry=is a general name for a group of biological methods used to measure various parameters of cells.
- 3. 1930s 1950s Counting of RBCs through capillary tube Development of coulter principle Development of FACS and other advances 1970s
- 5. Components of Flow Cytometry Fluidics Optics Electronics
- 6. The fluidics system transports particles in a stream to the laser beam for interrogation. Objective is to have one cell pass through the laser intercept at a time.
- 7. Fluorescent and SSC signals are collected at right angles to the excitation laser are progressively picked off to facilitate multiple fluorochrome use. The optics system consists of lasers to illuminate the particles in the sample stream and optical filters to direct the resulting light signals to the appropriate detectors.
- 8. The fluorescence intensity measured is proportional to the number of fluorescent molecules bound to the cell. For some instruments equipped with a sorting feature, the electronics system is also capable of initiating sorting decisions to charge and deflect particles. The electronics system converts the detected light signals into electronic signals that can be processed by the computer.
- 9. Scatter Signals and Fluorescent Signals Incident light scattered at small angles (0.5-2.0°) is called Forward Scatter (FSC) Incident light scattered at an angle of 90° is called Side Scatter (SSC) Scatter — Forward Scatter (FSC) — Side Scatter (SSC) Fluorescence — FITC, PE, APC, GFP, DAPI
- 10. Forward Scatter (FSC) Rough measure of size, influenced by the wavelength of light, and the angle, lenses and apertures that light is collected at and with. Different flow cytometers will give slightly different FSC measurements. Most flow cytometers measure FSC with a photodiode. Bacteria – Photomultiplier tube (PMT) Dead cells may have lower FSC measurements than live cells. Osmotic swelling can increase cell volume, and decrease light scatter. Scatter Signals and Fluorescent Signals
- 11. Side Scatter (SSC) SSC is the measure of light scattered at an angle of 90º (orthogonal). SSC is a measure of the complexity of the cell’s internal structures. The more ‘granular’ a cell is the higher its SSC will be. A neutrophil is much more granular than a lymphocyte. Scatter Signals and Fluorescent Signals
- 12. FSC has some similarities to size SSC has some similarities to granularity and complexity Scatter Signals and Fluorescent Signals
- 13. Fluorochromes Fluorochromes are substances that can be excited by certain light source (such as laser) and emit a fluorescent signal at a single wavelength. Fluorescent dyes can directly bind to certain cellular content, such as DNA and RNA, and allow us to perform quantitative analysis on individual cells. However, in most cases fluorochromes are conjugated with monoclonal antibodies, which specifically target cellular antigens/markers. Synthetic, organic dyes Proteins FITC PE Cy dyes APC Horizon dyes Green fluorescent protein eFluor dyes Pacific blue Brilliant violet
- 14. How to Choose Fluorochromes Fluorochromes Excitation (nm) Emission (nm) FITC 488 525 PE 488 575 PI 488 630 Cy5 488 675 PerCP 488 675 APC 633 660 APC-Cy 633 767 Antibody availability Function — i.e. Mcherry VS GFP Fluorochrome brightness Excitation source Emission filters Other fluorochromes
- 15. Visualizing Flow Cytometry Data Dot plot: one parameter vs another Density plot: viewing the frequency of subpopulations Contour plot: showing the probability contouring Histogram plot: one parameter only
- 16. USA Europe Tel: 1-631-626-9181 Fax: 1-631-614-7828 Email: info@creative-bioarray.com 45-1 Ramsey Road, Shirley, NY 11967, USA Tel: 44-207-097-1828 Email: info@creative-bioarray.com www.creative-bioarray.com