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FRET Analysis of Protein-Protein Interaction and Redox Sensor Folding Using the ImageStream

Merck
28 Jan 2011

This application note shows FRET measurement with different fluorphore combinations and experimental applications using the ImageStream imaging cytometer, which enables quantitative FRET measurement directly from images of a large numbers of cells per sample.

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Image Stream®X Mark II Imaging Flow Cytometer

Merck

The revolutionary ImageStream®X Mark II Imaging Flow Cytometer combines the speed, sensitivity, and phenotyping abilities of flow cytometry with the detailed imagery and functional insights of microscopy. This unique combination enables a broad range of applications that would be impossible using either technique alone. This instrument produces multiple high-resolution images of every cell directly in flow, including brightfield and darkfield (SSC), and up to 10 fluorescent markers with sensitivity exceeding conventional flow cytometers. Compared to its predecessor, the new ImageStream®X Mark II Imaging Flow Cytometer offers a streamlined workflow, greater flexibility, and optimizations for rare cell applications. Taken together, the capabilities of the ImageStream®X Mark II make it superior for traditional flow applications while greatly expanding the scope of flow cytometry. Applications include the study of cell-cell interactions, phagocytosis, apoptosis and autophagy, the characterization of circulating tumor cells, and many others.

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Fluorescence SpectroscopyFluorometers and spectrofluorometers (also called fluorescence spectrometers) are used to measure the intensity and wavelength of fluorescent light emitted from a sample after excitation by illumination. Spectrofluorometers utilize monochromators to select the desired wavelengths, whereas filter fluorometers employ a set of filters. Spectrofluorometers for measuring steady-state fluorescence and lifetime fluorescence (or time-resolved fluorescence) are available, as well as fluorescence microscopes and microplate readers. Find the best fluorescence spectroscopy products in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Cell / Tissue CultureCell culture or tissue culture is used to study the biology of cells or tissues and to isolate cellular products in an environment which can be manipulated and well defined. Accurately control your culture environment with bioreactors or culture incubators, bind your cells to a surface or together with an extracellular matrix. Distinguish cell types with differential media or proliferate cells with certain characteristics using selective media. Enrich your media with supplements such as growth factors, sera and vitamins. Find the best cell and tissue culture products, kits and equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.ProteomicsProteomics is the systemic bioinformatics study of proteins and amino acids, including their structure, size, function and identification. Tools used in proteomics include chromatography, blotting and gels, protein arrays, mass spectrometry and ELISA and associated analysis software. Analyzers and proteomic systems should be sensitive, high resolution, fast and may be automated for high-throughput.Flow Cytometry / Cell CountingFlow cytometers are used to count, sort and examine multiple characteristics of cells. Other cell analysis equipment includes image cytometers, cell counters, fluorescence-activated cell sorters (FACS), magnetic-activated cell sorters (MACS), and a range of flow cytometry assay kits. Flow cytometers can reveal information on cell viability, cell proliferation, apoptosis and cell cycle progression, as well as identify cell populations and intracellular or cell-surface molecules. Additionally, some flow cytometers, known as FACS, have an additional sorting function after analysis. Cell counters and image cytometers count live and dead cell populations and can also conduct cell proliferation assays. Find the best flow cytometers, cell counters and cell sorters in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.NanotechnologyNanotechnology, or nanotech, is an engineering technique using molecular scale functional systems. Applications of nanotechnology include medicine and medical devices, electronics, air and water purification, food science and energy production.Particle CharacterizationParticle characterization instruments are used to determine particle size distribution, shape, surface area, zeta potential, density and porosity of particles and materials. Multiple tecchniques are available for determining particle size, shape and count including dynamic light scattering (DLS), laser diffraction, electrozone (Coulter technique), imaging particle analysis and single particle optical sensing. Determine the density of your material with a gas pycnometer or examine its surface area and porosity with gas adsorption analyzers and mercury porosimeters. Find the best particle characterization instruments in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Light MicroscopyLight microscopes or optical microscopes are used to visualize microscale objects under magnification, including cells, clinical specimens and materials. Lab equipment for light microscopy includes confocal microscopes, fluorescence microscopes, zoom and stereo microscopes. Microscope slides and imaging reagents are available for visualizing samples, as well as various microscope stages and incubators for large or temperature-sensitive samples. Find the best light microscopes in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.

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FRET Analysis of Protein-Protein Interaction and Redox Sensor Folding Using the ImageStream