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PubMed · 3967553

Helpful hints in flow cytometry and sorting.

P N Dean
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Abstract

Most users of flow cytometers and sorters become proficient in their use through direct operation of the instruments. There are no text books or operation manuals that describe all of the problems that will be encountered and that may not be apparent. We have discovered many of these problems through finding an explanation for unexpected data and in resolving difficulties in achieving and maintaining high resolution and stable operation of the instruments. This paper presents a summary of problems encountered and solutions found at this laboratory and others.

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FiltrationFlow CytometryFluorescenceLasersComparative StudyJournal ArticleResearch Support, U.S. Gov't, Non-P.H.S.

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P N Dean. 1985. Helpful hints in flow cytometry and sorting.. https://doi.org/10.1002/cyto.990060111

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Analog filtering of large solvent signals for improved dynamic range in high-resolution NMR.

The large solvent signal from samples in H2O solvent still challenges the dynamic range capability of any spectrometer. The solvent signal can be largely removed with a pair of simple resistor-capacitor (RC) high-pass filters when the solvent frequency is set at center band (zero frequency) using quadrature detection, with RC approximately 0.5 ms. However, an approximately 0.5-ms transient remains at initial time, which we reduce fourfold for a short time only, just before the A/D converter, by means of a variable-gain amplifier, and later restore with software. This modification can result in a nearly fourfold increase in dynamic range. When we converted to a frequency-shifted mode (A. G. Redfield and S. D. Kunz, 1994, J. Magn. Reson. A 108, 234-237) we replaced the RC high-pass filter with a quadrature feedback notch filter tuned to the solvent frequency (5.06 kHz). This filter is an example of a class of two-input/two-output filters which maintain the spectral integrity (image-free character) of quadrature signals. Digital filters of the same type are also considered briefly. We discuss the implications of these ideas for spectrometer input design, including schemes for elimination of radiation damping, and effects of probe bandwidth on extreme oversampling.

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