Optical analysis of the AMAC IIIS transducer

Leif, Robert C.; Wells, Mark

doi:10.1364/ao.26.003244

Appl. Opt.

1987

DOI: 10.1364/ao.26.003244

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Optical analysis of the AMAC IIIS transducer

Robert C. Leif¹,

Mark Wells²

Abstract: The AMAC IIIS electrooptical transducer for flow cytometry facilitates simultaneous measurements of electrical impedance and several optical properties of single cells. Ray-trace analysis demonstrates that its spherical exterior permits light rays to emerge effectively undeviated and thus lowers the required numerical aperture of the collecting lens as compared with a flat surfaced transducer. This feature results in an increased light-gathering ability, an increased depth of focus, minimum spherical and chrom… Show more

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Cited by 3 publications

References 6 publications

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Increasing sensitivity of the Ortho analytical cytofluorograph by modifying the fluid system

Zucker¹,

Elstein²,

Gershey

et al. 1990

Cytometry

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By adding Teflon tubing to the effluent line of the Ortho 50H analytical cytofluorograph, we were able to increase the sensitivity of fluorescence and scatter detection threefold without compromising resolution. By increasing sheath backpressure (resistance), the additional tubing increases particle residence time within the detection area and thereby increases the total photon emission density per particle. In addition, the longer, For maximum flow cytometer sensitivity and resolution, particles must be analyzed within a quartz flow cell. To achieve this, the sample stream is focussed hydrodynamically through a square flow cell orifice, thereby eliminating the optical aberrations associated with curved surfaces (1,3). High-resolution optics and additional lenses have also been employed to maximize the focussing of scattered and emitted light onto sensing devices (2,4-6). However, given these optical and fluidic considerations, little attention has been paid to the effect of effluent tubing dimensions on analytical flow cell hydrodynamics.Typically, the effluent stream is slowed to a n acceptable velocity by adding approximately one foot of 0.3-mm inner diameter (i.d.1 tubing suspended over a waste reservoir. However, this tubing is often the site of blockages that result in signal-altering turbulence.To correct this, users cut off the clogged ends thereby shortening the coil and, consequently, increasing the flow rate. Eventually, particle residence time becomes too short to produce acceptable signals and the coil is replaced.To assess the effect of effluent tubing dimensions on sensitivity, we measured the integrated green fluorescence and scatter signals of fluorescent polystyrene beads as a function of length of 0.30-mm) 0.59-mm or 0.81-mm i.d. effluent tubing. Our results indicate that both the sensitivity of detection and system stability can be improved by altering effluent tubing length andlor inner diameter.wider tubing dampens the stream-destabilking effects of minor obstructions in or movement of the effluent line. This increase in sensitivity and stability is desirable in applications requiring detection of particles exhibiting low-level fluorescence.

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Increasing sensitivity of the Ortho analytical cytofluorograph by modifying the fluid system

Zucker¹,

Elstein²,

Gershey

et al. 1990

Cytometry

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show abstract

Use of a spherical multiparameter transducer for flow cytometry

Leif¹,

Cayer²,

Dailey³

et al. 1995

Cytometry

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Fused silicon dioxide, multiparameter flow transducers with 50 pm internal square cross section and -60 pm length can simultaneously measure DC and RF impedance as well as fluorescence and multiple-angle light scattering. A spherical version of such a transducer was mounted in an EPICS CVA flow-cell housing and was installed on a research prototype equipped with an argon-ion laser. The signal that was produced by the spherical transducer with EPICS DNA-Check beads w a s 1.73 times greater than that produced with the standard cylindrical flow cell. Similarly, with EPICS Immuno-Brite beads, the average ratio was 1.96.The Coulter impedance and light-scattering measurements were similar to those produced with the conventional cylindrical outside flow cell, although the internal cross section of the sphere was square and that of the cylinder was circular. The theoretical arguments of Leif and Wells have been demonstrated to be correct. At present, monolithic, spherical fused-silica transducers are the optimal design for combined electrooptical, multiparameter flow cytometry analyzers. o 1995 WUey-Liss, Inc.

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Trends and Developments in Flow Cytometry Instrumentation

Shapiro

1993

Annals of the New York Academy of Sciences

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Optical analysis of the AMAC IIIS transducer

Cited by 3 publications

References 6 publications

Increasing sensitivity of the Ortho analytical cytofluorograph by modifying the fluid system

Increasing sensitivity of the Ortho analytical cytofluorograph by modifying the fluid system

Use of a spherical multiparameter transducer for flow cytometry

Trends and Developments in Flow Cytometry Instrumentation

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