Nozzle design parameters and their effects on rapid sample delivery in flow cytometry

Graves, Steven W.; Nolan, John P.; Jett, James H.; Martin, John; Sklar, Larry A.

doi:10.1002/cyto.10056

Cited by 29 publications

(31 citation statements)

References 18 publications

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“…As a result, a transient widening of the sheath stream occurs. The optical components associated with the detection of light scatter and fluorescence are sensitive to the width of the stream (6,11). However, the effect of sample stream widening can be mitigated by the use of light scatter gates (Fig.…”

Section: Results and Discussion Characteristics Of The Rapid MIXmentioning

confidence: 99%

“…Each fluidic part was designed to be interchangeable to meet the requirements of desired experimental conditions. A rapid mixing device for sample delivery to a Becton-Dickinson FACScan flow cytometer (Becton-Dickinson Immunocytometry System, San Jose, CA, USA) was developed using commercial components (1,6,7). The core component of the rapid-mixing device, FIA 3000, consisted of a control module with three computer-driven syringes (Cavro Scientific Instruments, Sunnyvale, CA, USA), two syringe pumps, two three-way isolated solenoid valves (NResearch, West Caldwell, NJ, USA), and FIALab software (FIALab Instruments, Bellevue, WA, USA) to control the flow of rapidly mixed samples.…”

Section: Rapid-mixing Componentsmentioning

confidence: 99%

“…For the past decade, one of us has been involved in the development of the mixing method of continuous analysis, which integrates a computer-controlled and syringebased mixing device with a flow cytometer (1)(2)(3)(6)(7)(8)(9)(10). A prevailing technical issue of concern in this work involves the orthogonal fluidic requirements of rapid-mix devices and the flow cytometers.…”

mentioning

confidence: 99%

“…It is therefore important that the introduction of the sample into the sheath flow must be under laminar flow conditions to conserve the hydrodynamic stability of the sheath fluid. One approach was to use a series combination of syringes, valves, and nozzle designs and pressure regulation to achieve rapid stability of the sheath fluid after delivery of sample from a rapid mix device (1,6). This approach has worked well for relatively large-volume assays (milliliter scale) and is generally not applicable to small-volume assays.…”

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confidence: 99%

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Small‐volume rapid‐mix device for subsecond kinetic analysis in flow cytometry

Zwartz

López

et al. 2005

Cytometry Pt A

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Background: Rapid-mix flow cytometry has emerged as a powerful tool for mechanistic analysis of ligand binding, cell response, and molecular assembly. Although progress has come from improving sample delivery capabilities, little attention has been paid to the volumetric requirements associated with precious biological reagents. Methods: By using programmable syringes, valves, and other fluidic components, we created a modular, precisely regulated rapid-mix device for the delivery of smallvolume samples to the flow cytometer. The device was tested using a bead-based assay in which the binding kinetics between native biotin and fluorescein biotin-bearing beads were characterized. Results: Bead suspensions and reagents paired in 35-to 45-ll aliquots were efficiently mixed by the device and

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Section: Results and Discussion Characteristics Of The Rapid MIXmentioning

confidence: 99%

Section: Rapid-mixing Componentsmentioning

confidence: 99%

mentioning

confidence: 99%

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confidence: 99%

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Small‐volume rapid‐mix device for subsecond kinetic analysis in flow cytometry

Zwartz

López

et al. 2005

Cytometry Pt A

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“…While fluorescent formyl peptides and FPR-GFP made it possible to study ternary complex dynamics in real-time, sample mixing and delivery times in flow cytometry limited kinetic resolution [16]. To minimize the dead time, online mixing schemes have been developed to achieve subsecond resolution [17]. However, for ternary complex reagents available only in limited quantities, we have recently described a small volume rapid mixing device [18] to measure subsecond disassembly kinetics in the presence of saturating concentrations of guanine nucleotides.…”

Section: Introductionmentioning

confidence: 99%

Rapid-mix flow cytometry measurements of subsecond regulation of G protein-coupled receptor ternary complex dynamics by guanine nucleotides

Buranda

Simons

et al. 2007

Analytical Biochemistry

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We have used rapid mix flow cytometry to analyze the early subsecond dynamics of the disassembly of ternary complexes of G-protein coupled receptors (GPCRs) immobilized on beads to examine individual steps associated with guanine nucleotide activation. Our earlier studies suggested that the slow dissociation of Gα and Gβγ subunits was unlikely to be an essential component of cell activation. However, these studies did not have adequate time resolution to define precisely the disassembly kinetics. Ternary complexes were assembled using three formyl peptide receptor constructs (wild type, FPR-Gα i2 fusion, and FPR-GFP fusion) and two isotypes of the α subunit (α i2 and α i3 ) and βγ dimer (β 1 γ 2 and β 4 γ 2 ). At saturating nucleotide levels, the disassembly of a significant fraction of ternary complexes occurred on a subsecond time frame for α i2 complexes and τ 1/2 ≤ 4 sec for α i3 complexes, time scales which are compatible with cell activation. β 1 γ 2 isotype complexes were generally more stable than β 4 γ 2 associated complexes. The comparison of the three constructs however proved that the fast step was associated with the separation of receptor and G protein and that the dissociation of the ligand or of the α and βγ subunits was slower. These results are compatible with a cell activation model involving G protein conformational changes rather than disassembly of Gαβγ heterotrimer.

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Fluidics

Suthanthiraraj

Graves

2013

CP Cytometry

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The use of fluidics is implicit in a technology named “flow cytometry”, which flows a cell or particle through a sensing volume to obtain serial analysis of particles on a one by one basis. This flow of particles enables flow cytometry to collect information on multiple particle populations, giving it a distinct advantage over bulk analysis approaches. Moreover, flow cytometers can analyze thousands of particles per second in a single flowing stream. Additionally, use of volumetric sample delivery makes it possible for flow cytometers to accurately count cells and particles. Furthermore, the analysis results can be coupled with a fluidic diversion mechanism to sort and collect particles based on desired properties. Finally, when high throughput sampling technologies are employed to rapidly change the input of the sample stream, a flow cytometer can become an integral tool for high throughput screening. The above properties have made flow cytometry useful in a wide range of biomedical applications. In this unit we will present an overview of fluidic systems that make flow cytometry possible. This will introduce historical approaches, explanations of the commonly implemented current fluidics, and brief discussions of potential future fluidics where appropriate.

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Nozzle design parameters and their effects on rapid sample delivery in flow cytometry

Cited by 29 publications

References 18 publications

Small‐volume rapid‐mix device for subsecond kinetic analysis in flow cytometry

Small‐volume rapid‐mix device for subsecond kinetic analysis in flow cytometry

Rapid-mix flow cytometry measurements of subsecond regulation of G protein-coupled receptor ternary complex dynamics by guanine nucleotides

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