cytometry Protocols

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Flow Cytometry Data Analysis Protocols

Flow Cytometry Analysis Protocol. Springer Lab - Harvard. Background on flow cytometry technique: Flow cytometry is a method which uses instrumentation scanning single cells flowing past excitation sources in a liquid medium.
- [Read Flow Cytometry Analysis Protocol]

Category: Cell Biology and Cell Culture: Apoptosis Protocols

Flow Cytometry Laboratory. Apoptosis. Perdue. - [Read Flow Cytometry Laboratory]

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Apoptosis Cytometry Protocols

Common methods applicable to flow cytometry make it possible to: (1) identify and quantify dead or dying cells, (2) reveal a mode of cell death (apoptosis or necrosis), and (3) study mechanisms involved in cell death. Gross changes in cell morphology and chromatin condensation, which occur during apoptosis, can be detected by analysis with laser light beam scattering. - [Read Flow Cytometry of Apoptosis Protocol]

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Flow Cytometry Immunofluorescence Staining Protocols

Protocol provides a method for analysis of cell surface antigens using direct immunofluorescence, i.e., when the antibody being used is directly labeled to a fluorescent dye. After staining, the cells can be analyzed immediately by flow cytometry or fixed and stored for later analysis. - [Read Fluorescent Staining of Cell Surface Antigens on Viable Cells Using Direct Immunofluorescence]

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Flow Cytometry Immunofluorescence Staining Protocols

Protocol for immunodetection of cyclin D1 and D2/D3 using 488/630 nm dual laser flow cytometry. - [Read Immunodetection of cyclin D1 and D2/D3 using 488/630 nm dual laser flow cytometry Protocol]

Category: Immunology: Immunofluorescence Protocols: Immunofluorescence Staining Protocols

Protocols for immunofluorescent staining for flow cytometry. Includes: Protocol for Staining of Cell Suspensions of Murine Lymphoid Tissue; Protocol for Staining of Human Peripheral Blood; Fluorescent Dyes for Flow Cytometric Analysis - [Read Immunofluorescent Staining for Flow Cytometry Protocols]

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols

Protocol for in vitro class switching staining for flow cytometry. - [Read In Vitro Class Switching Staining for Flow Cytometry Protocol]

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols

An Indirect Immunofluorescence Labeling Protocol for flow cytometry. Iowa State University, Flow Cytometry Facility - [Read Indirect Immunofluorescence Labeling Protocol]

Category: Signalling Signal Transduction Protocols: Cytokine Protocols

Intracellular Cytokine Staining Protocol. Analysis of surface molecules and intracellular cytokines at the single-cell level using FACS or flow cytometry. - [Read Intracellular Staining for Flow Cytometric Analysis Protocols]

Category: Immunology: Mononuclear Cell Isolation Protocols: B Lymphocyte Isolation Protocols

A flow cytometry technique is presented, which results in the selection and isolation of two populations of cells from a complex mixture based on physical properties (e.g., size and internal granularity) and correlated expression of several surface molecules - [Read Isolation of Ly-1+/CD5+ B Cells by Cell Sorting Protocol]

Category: Cell Biology and Cell Culture: Cell Viability Protocols

Assay measures cell viability. It is a two-color fluorescence assay that simultaneously determines Live cell number and Dead cell number. This protocol is designed for use with the GEMINI XS Microplate Spectrofluorometer, a multi-well plate scanner with dual excitation/emission capabilities, but the assay is also adaptable for flow cytometry and fluorescence microscopy. Includes: Cell Culture; Preparation for the Assay; Live/Dead Assay; Reading the Plate; Data Analysis; Alternative protocol. - [Read Live/Dead Assay for Cell Viability Protoco]

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: DNA Staining Protocols for Flow Cytometry

This protocol describes a method for quantitative measurement of DNA using propidium iodide (PI) staining and flow cytometry. PI stains all double-stranded regions of both DNA and RNA by intercalating between the stacked bases of the double helix. PI cannot penetrate an intact cell membrane; therefore, cells are fixed prior to staining. The ethanol-fixed cells can be stored unstained at 4°C for days, or even weeks, and then stained and analyzed. - [Read Measurement of DNA Content Using Propidium Iodide (PI) Staining of Fixed Whole Cells Protocol]

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols

The more commonly available single-laser cytometers can also be used to measure multicellular conjugates, but due to overlaps in emission spectra, the extent of labeling cells with fluorophores must be controlled much more carefully when the single-laser machines are used. This protocol describes the labeling of cells and analysis of conjugates with either dual-laser or single laser flow cytometers. - [Read Measurement of Intercellular Conjugates by Flow Cytometry Protocol]

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Intracellular Ion Measurement Protocols

Describes flow cytometric protocols using the dyes Indo-1 AM, Fluo-3, and Fura Red AM to measure intracellular calcium concentration. Support protocols detail the use of calcium buffers to calibrate a flow cytometric calcium assay, and methods to facilitate dye loading; an alternate protocol describes the use of a spectrofluorimeter to measure intracellular calcium for those investigators without access to a flow cytometer. - [Read Measurement of Intracellular Ions by Flow Cytometry Protocol]

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Apoptosis Cytometry Protocols

Protocol for measuring apoptosis and necrosis by dual laser flow cytometry. Includes: Preparation of stock solutions of HO342, PI, and 7-AAD; Staining for detection of apoptosis; - [Read Measuring Apoptosis and Necrosis by Dual Laser Flow Cytometry Protocol]

Category: Cell Biology and Cell Culture: Apoptosis Protocols

Measuring Apoptosis and Necrosis (Oncosis) by Dual Layer Flow Cytometry - [Read Measuring Apoptosis and Necrosis by Dual Layer Flow Cytometry]

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols

Protocol for membrane potential analysis by flow cytometry. Includes: Preparation of the dyes; Loading of the dyes; Flow Cytometric Analysis. - [Read Membrane Potential Analysis by Flow Cytometry Protoocol]

Category: Molecular Imaging: Immunofluorescence Microscopy

Minimizing Background in Immunofluorescence Procedures. Includes tips and methods on reducing background staining. Janis V. Giorgi Flow Cytometry Laboratory, UCLA. - [Read Minimizing Background in Immunofluorescence Procedures]

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols

Provides an overview of terminology and echniques unique to flow ytometry and is divided into two sections. The first section discusses technical aspects of flow cytometry which apply primarily to the instrumentation-oriented flow cytometry phase. The second section discusses electronic cell separation using flow cytometry. - [Read Overview of Flow Cytometry]

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Flow Cytometry Immunofluorescence Staining Protocols

Protocol for phenotype-specific immunodetection of cyclins using 488/630 nm dual laser flow cytometry. This protocol is for use with the D and E cyclins and employs 488 nm argon laser excitation of propidium iodide and a FITC-conjugated phenotypic label, and 630 nm NeNe or diode laser excitation of the fluorochrome Cy5 to detect cell cycle-specific cyclin D expression. - [Read Phenotype-Specific Immunodetection of Cyclins using 488/630 nm Dual Laser Flow Cytometry Protocol]

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols

A procedure for direct and indirect staining of single-cell suspensions of lymphoid tissue or peripheral blood lymphocytes to detect cell surface membrane antigens is presented. In addition, support protocols present methods for fluorescence labeling of purified antibodies. A protocol for flow cytometric analysis of intracellular antigens in single-cell suspensions is also included. - [Read Preparation of Cells and Reagents for Flow Cytometry Protocols]

Category: Yeast Protocols: Yeast Nucleic Acid Protocols

Flow cytometry is used to analyze the quantity of DNA in cells. Since the DNA content of cells varies through the cell cycle, this information can provide an indication of cell cycle progression. This protocol uses SYTOX Green staining. - [Read Preparation of Yeast Cells for Flow Cytometry Protocol]

Category: Immunology: T Cell Protocols

T cell hybridomas can be obtained by fusing activated T cells with tumor cells. A heterogeneous population of hybridomas can be cloned by limiting dilution to obtain hybridomas that express specificity to one T cell receptor (TCR). Protocol describes cell fusion and selection of T cell hybridomas. A protocol is provided for screening of T cell hybridomas for expression of the CD3-TCR complex by flow cytometry analysis. - [Read Production of Mouse T Cell Hybridomas Protocol]

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: DNA Staining Protocols for Flow Cytometry

This protocol describes a method for quantitative measurement of DNA in solid tissue samples using either propidium iodide (PI) or DAPI staining followed by flow cytometry. PI can be excited at 488 nm by the argon-ion laser, the most commonly used laser in flow cytometry. Alternatively, DAPI is best excited by a high-power UV laser, which is less commonly available. - [Read Propidium Iodide (PI) or DAPI Staining of Unfixed Solid Tissues for Flow Cytometry Protocol]

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: DNA Staining Protocols for Flow Cytometry

This protocol describes a method for quantitative measurement of DNA in tissue culture cells using either propidium iodide (PI) or DAPI staining followed by flow cytometry. PI can be excited at 488 nm by the argon-ion laser, the most commonly used laser in flow cytometry. Alternatively, DAPI is best excited by a high-power UV laser, which is less commonly available. - [Read Propidium Iodide (PI) or DAPI Staining of Unfixed Tissue Culture Cells for Flow Cytometry Protocol]