No-wash, No-lyse Detection of Phagocytic Cells via a pHrodo BioParticles Functional Assay in Human Whole Blood on the Attune NxT Flow Cytometer

No-wash, No-lyse Detection of Phagocytic Cells via a pHrodo BioParticles Functional Assay in Human Whole Blood on the Attune NxT Flow Cytometer

18 Oct 2015

Analysis of biological samples in the most physiologically relevant state with minimal sample preparation and manipulation is a key objective in many cell biology workflows. Normally, whole blood samples require time-consuming enrichment and manipulation in preparation for analysis on conventional hydrodynamic focusing flow cytometers. These manipulations can result in loss of rare cell types and undesirable phenotypic changes. This application note outlines a strategy for a no-wash, no-lyse approach to a functional assay using Molecular Probes™ pHrodo™ BioParticles™ Conjugates and the Applied Biosystems™ Attune™ NxT Flow Cytometer to assess phagocyte function in human whole blood.

Invitrogen™ Attune™ NxT Flow Cytometer

Thermo Fisher Scientific

The Invitrogen™ Attune™ NxT flow cytometer can be designed to accommodate the most common fluorophores and fluorescent proteins used in flow cytometry to match the panels you are currently running. Multiple fluorescent proteins can be detected with an optional 561 nm laser. Whether you configure your system now or upgrade later, the Attune NxT flow cytometer can grow with you and your research needs. The Attune NxT flow cytometer, designed using acoustic-assisted hydrodynamic focusing, achieves sample-throughput rates of of 12.5 µL/min–1 mL/min or 12.5–1,000 µL/min—up to 10 times faster than traditional hydrodynamic focusing systems and data acquisition speeds of 35,000 events/sec. Acoustic focusing allows significantly higher sample rates, which simplifies sample preparation, reduces protocol times, and minimizes cell loss. This feature is useful for no wash, no lyse immunophenotyping protocols using whole blood and for samples that are inherently low in concentration, including cerebrospinal fluid and stem cell preparations. Your next result relies on the ability of your experimental design and your instrument to deliver more—more data, more detail, and more throughput. The Attune NxT flow cytometer was designed to meet the demands of more complex experiments, offering reliable data, day after day, in a grow-with-you system.   Features & Benefits: Modular design—Multiple configurations available—field upgradable. Save time—10X faster speeds with no loss in data quality. Simplified sample prep—No wash, no lyse options, non-clogging fluidics. Enables unique applications—Complex protocols on a broad range of cell types and samples.

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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.ImmunologyImmunological techniques measure and characterize immune responses. Immunology kits and analysis systems often use techniques such as ELISA, radioimmunoassay (RIA) and immunodiffusion assays, Immunohistochemistry, and flow cytometry. Immunologists use equipment such as flow Cytometers, plate readers, plate washers and fluorescent microscopes.Cell-Based AssaysCell-based assays are used to monitor the presence, quantity and activities of a desired cellular analyte including drug molecules or biomarkers. This can reveal information on cell health (apoptosis, cytotoxicity, viability and proliferation assays), cell metabolism, cell migration and cell signaling mechanisms. Find the best cell-based assay products, kits and equipment with our peer reviewed product directory: compare products, check customer reviews and receiving pricing direct from manufacturers.Cell ManipulationCell WashingCell BiologyCell biology studies the structure, function, and behavior of cells. Understanding cellular processes is essential for research in areas such as cancer biology, stem cell research, and neurobiology. Techniques like flow cytometry, microscopy, and cell culture enable researchers to explore cellular mechanisms in detail. Browse our peer-reviewed product directory to find the best cell biology tools and equipment, compare products, check customer reviews, and get pricing directly from manufacturers.Phenotypic ScreeningPhenotypic screening assesses cellular responses to compounds, enabling drug discovery and target identification. This technique is pivotal in finding effective treatments for complex diseases. Discover phenotypic screening platforms and tools with peer-reviewed comparisons and pricing in our directory.Lysis
No-wash, No-lyse Detection of Phagocytic Cells via a pHrodo BioParticles Functional Assay in Human Whole Blood on the Attune NxT Flow Cytometer