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